Kinase inhibitors

ABSTRACT

There are provided compounds of formula I, 
     
       
         
         
             
             
         
       
     
     wherein:
 
Y represents NR 2 R 3 ;
 
one of R 2  and R 3  represents —[C 2-4  alkylene-O] 1-12 —[C 2-4  alkylene]-R 2a  and the other of R 2  and R 3  has a meaning given in the description; and
 
R, R 1 , R 2a , R a , R b , Q, X and Y have meanings given in the description,
 
which compounds have antiinflammatory activity (e.g., through inhibition of one or more of members of: the family of p38 mitogen-activated protein kinase enzymes; Syk kinase; and members of the Src family of tyrosine kinases) and have use in therapy, including in pharmaceutical combinations, especially in the treatment of inflammatory diseases, including inflammatory diseases of the lung, eye and intestines.

FIELD OF THE INVENTION

The invention relates to compounds which are inhibitors of the family ofp38 mitogen-activated protein kinase enzymes (referred to herein as p38MAP kinase inhibitors), for example the alpha and gamma sub-typesthereof, and of Syk kinase and the Src family of tyrosine kinases, andto their use in therapy, including in pharmaceutical combinations,especially in the treatment of inflammatory diseases, in particularinflammatory diseases of the lung, such as asthma and COPD, as well asthose of the gastrointestinal tract, such as ulcerative colitis andCrohn's disease, and of the eye, such as uveitis.

BACKGROUND OF THE INVENTION

The listing or discussion of an apparently prior-published document inthis specification should not necessarily be taken as an acknowledgementthat the document is part of the state of the art or is common generalknowledge.

Four p38 MAPK isoforms (alpha, beta, gamma and delta respectively), eachdisplaying different patterns of tissue expression, have beenidentified. The p38 MAPK alpha and beta isoforms are found ubiquitouslyin the body, being present in many different cell types. The alphaisoform is well characterized in terms of its role in inflammation.Although studies using a chemical genetic approach in mice indicate thatthe p38 MAPK beta isoform does not play a role in inflammation (O'Keefe,S. J. et al., J Biol Chem., 2007, 282(48):34663-71), it may be involvedin pain mechanisms through the regulation of COX2 expression(Fitzsimmons, B. L. et al., Neuroreport, 2010, 21(4):313-7). Theseisoforms are inhibited by a number of previously described smallmolecular weight compounds. Early classes of inhibitors were highlytoxic due to the broad tissue distribution of these isoforms whichresulted in multiple off-target effects of the compounds. Furthermore,development of a substantial number of inhibitors has been discontinueddue to unacceptable safety profiles in clinical studies (Pettus, L. H.and Wurz, R. P., Curr. Top. Med. Chem., 2008, 8(16):1452-67). As theseadverse effects vary with chemotype, and each of these compounds hasdistinct kinase selectivity patterns, the toxicities observed may bestructure—rather than p38 mechanism-based.

Less is known about the p38 MAPK gamma and delta isoforms, which, unlikethe alpha and beta isozymes are expressed in specific tissues and cells.The p38 MAPK-delta isoform is expressed more highly in the pancreas,testes, lung, small intestine and the kidney. It is also abundant inmacrophages and detectable in neutrophils, CD4+ T cells and inendothelial cells (Shmueli, O. et al., Comptes Rendus Biologies, 2003,326(10-11):1067-1072; Smith, S. J. Br. J. Pharmacol., 2006, 149:393-404;Hale, K. K., J. Immunol., 1999, 162(7):4246-52; Wang, X. S. et al., J.Biol. Chem., 1997, 272(38):23668-23674.) Very little is known about thedistribution of p38 MAPK gamma although it is expressed more highly inbrain, skeletal muscle and heart, as well as in lymphocytes andmacrophages. (Shmueli, O. et al., Comptes Rendus Biologies, 2003,326(10-11):1067-1072, (2003)/; Hale, K. K., J. Immunol., 1999,162(7):4246-52: Court, N. W. et al., J. Mol. Cell. Cardiol., 2002,34(4):413-26; Mertens, S. et al., FEBS Lett., 1996, 383(3):273-6.)

Selective small molecule inhibitors of p38 MAPK gamma and p38 MAPK deltaare not currently available, although one previously disclosed compound,BIRB 796, is known to possess pan-isoform inhibitory activity. Theinhibition of p38 MAPK gamma and delta isoforms is observed at higherconcentrations of the compound than those required to inhibit p38 MAPKalpha and p38 beta (Kuma, Y. J. Biol. Chem., 2005, 280:19472-19479). Inaddition BIRB 796 also impaired the phosphorylation of p38 MAPKs or JNKsby the upstream kinase MKK6 or MKK4. Kuma discussed the possibility thatthe conformational change caused by the binding of the inhibitor to theMAPK protein may affect the structure of both its phosphorylation siteand the docking site for the upstream activator, thereby impairing thephosphorylation of p38 MAPKs or JNKs.

p38 MAP kinase is believed to play a pivotal role in many of thesignalling pathways that are involved in initiating and maintainingchronic, persistent inflammation in human disease, for example, insevere asthma, COPD (Chung, F., Chest, 2011, 139(6):1470-1479) andinflammatory bowel disease (IBD). There is now an abundant literaturewhich demonstrates that p38 MAP kinase is activated by a range ofpro-inflammatory cytokines and that its activation results in therecruitment and release of additional pro-inflammatory cytokines.Indeed, data from some clinical studies demonstrate beneficial changesin disease activity in patients during treatment with p38 MAP kinaseinhibitors. For instance Smith describes the inhibitory effect of p38MAP kinase inhibitors on TNFα (but not IL-8) release from human PBMCs.

The use of inhibitors of p38 MAP kinase in the treatment of COPD and IBDhas also been proposed. Small molecule inhibitors targeted to p38 MAPKα/β have proved to be effective in reducing various parameters ofinflammation in:

-   -   cells and tissues obtained from patients with COPD, who are        generally corticosteroid insensitive, (Smith, S. J., Br. J.        Pharmacol., 2006, 149:393-404);    -   biopsies from IBD patients (Docena, G. et al., J. of Trans.        Immunol., 2010, 162:108-115); and    -   in vivo animal models (Underwood, D. C. et al., Am. J. Physiol.,        2000, 279:L895-902; Nath, P. et al., Eur. J. Pharmacol., 2006,        544:160-167).

Irusen and colleagues also suggested the possibility of involvement ofp38 MAPKα/β on corticosteroid insensitivity via the reduction of bindingaffinity of the glucocorticoid receptor (GR) in nuclei (Irusen, E. etal., J. Allergy Clin. Immunol., 2002, 109:649-657). Clinical experiencewith a range of p38 MAP kinase inhibitors, including AMG548, BIRB 796,VX702, SCIO469 and SCIO323 has been described (Lee, M. R. and Dominguez,C., Current Med. Chem., 2005, 12:2979-2994). However, the major obstaclehindering the utility of p38 MAP kinase inhibitors in the treatment ofhuman chronic inflammatory diseases has been the toxicity observed inpatients. This has been sufficiently severe to result in the withdrawalfrom clinical development of many of the compounds progressed, includingall those specifically mentioned above.

COPD is a condition in which the underlying inflammation is reported tobe substantially resistant to the anti-inflammatory effects of inhaledcorticosteroids. Consequently, a superior strategy for treating COPDwould be to develop an intervention which has both inherentanti-inflammatory effects and the ability to increase the sensitivity ofthe lung tissues of COPD patients to inhaled corticosteroids. A recentpublication of Mercado (Mercado, N., et al., Mol. Pharmacol., 2011,80(6):1128-1135) demonstrates that silencing p38 MAPK gamma has thepotential to restore sensitivity to corticosteroids. Consequently theremay be a dual benefit for patients in the use of a p38 MAP kinaseinhibitor for the treatment of COPD and severe asthma.

Many patients diagnosed with asthma or with COPD continue to suffer fromuncontrolled symptoms and from exacerbations of their medical conditionthat can result in hospitalization. This occurs despite the use of themost advanced, currently available treatment regimens, comprising ofcombination products of an inhaled corticosteroid and a long actingβ-agonist. Data accumulated over the last decade indicates that afailure to manage effectively the underlying inflammatory component ofthe disease in the lung is the most likely reason that exacerbationsoccur. Given the established efficacy of corticosteroids asanti-inflammatory agents and, in particular, of inhaled corticosteroidsin the treatment of asthma, these findings have provoked intenseinvestigation. Resulting studies have identified that some environmentalinsults invoke corticosteroid-insensitive inflammatory changes inpatients' lungs. An example is the response arising fromvirally-mediated upper respiratory tract infections (URTI), which haveparticular significance in increasing morbidity associated with asthmaand COPD.

Epidemiologic investigations have revealed a strong association betweenviral infections of the upper respiratory tract and a substantialpercentage of the exacerbations suffered by patients already diagnosedwith chronic respiratory diseases. Some of the most compelling data inthis regard derives from longitudinal studies of children suffering fromasthma (Papadopoulos, N. G., Papi, A., Psarras, S. and Johnston, S. L.,Paediatr. Respir. Rev., 2004, 5(3):255-260). A variety of additionalstudies support the conclusion that a viral infection can precipitateexacerbations and increase disease severity. For example, experimentalclinical infections with rhinovirus have been reported to causebronchial hyper-responsiveness to histamine in asthmatics which isunresponsive to treatment with corticosteroids (Grunberg, K., Sharon, R.F., et al., Am. J. Respir. Crit. Care Med., 2001, 164(10):1816-1822).Further evidence derives from the association observed between diseaseexacerbations in patients with cystic fibrosis and HRV infections (Wat,D., Gelder, C., et al., J. Cyst. Fibros., 2008, 7:320-328). Alsoconsistent with this body of data is the finding that respiratory viralinfections, including rhinovirus, represent an independent risk factorthat correlates negatively with the 12 month survival rate in pediatric,lung transplant recipients (Liu, M., Worley, S., et al., Transpl.Infect. Dis., 2009, 11(4):304-312).

Clinical research indicates that the viral load is proportionate to theobserved symptoms and complications and, by implication, to the severityof inflammation. For example, following experimental rhinovirusinfection, lower respiratory tract symptoms and bronchialhyper-responsiveness correlated significantly with virus load (Message,S. D., Laza-Stanca, V., et al., PNAS, 2008; 105(36):13562-13567).Similarly, in the absence of other viral agents, rhinovirus infectionswere commonly associated with lower respiratory tract infections andwheezing, when the viral load was high in immunocompetent pediatricpatients (Gerna, G., Piralla, A., et al., J. Med. Virol., 2009,81(8):1498-1507).

Interestingly, it has been reported recently that prior exposure torhinovirus reduced the cytokine responses evoked by bacterial productsin human alveolar macrophages (Oliver, B. G., Lim, S., et al., Thorax,2008, 63:519-525). Additionally, infection of nasal epithelial cellswith rhinovirus has been documented to promote the adhesion of bacteria,including S. aureus and H. influenzae (Wang, J. H., Kwon, H. J. andYong, J. J., The Laryngoscope, 2009, 119(7):1406-1411). Such cellulareffects may contribute to the increased probability of patientssuffering a lower respiratory tract infection following an infection inthe upper respiratory tract. Accordingly, it is therapeutically relevantto focus on the ability of novel interventions to decrease viral load ina variety of in vitro systems, as a surrogate predictor of their benefitin a clinical setting.

High risk groups, for whom a rhinovirus infection in the upperrespiratory tract can lead to severe secondary complications, are notlimited to patients with chronic respiratory disease. They include, forexample, the immune compromised who are prone to lower respiratory tractinfection, as well as patients undergoing chemotherapy, who face acute,life-threatening fever. It has also been suggested that other chronicdiseases, such as diabetes, are associated with a compromisedimmuno-defense response. This increases both the likelihood of acquiringa respiratory tract infection and of being hospitalized as a result(Peleg, A. Y., Weerarathna, T., et al., Diabetes Metab. Res. Rev., 2007,23(1):3-13; Kornum, J. B., Reimar, W., et al., Diabetes Care, 2008,31(8): 1541-1545).

Whilst upper respiratory tract viral infections are a cause ofconsiderable morbidity and mortality in those patients with underlyingdisease or other risk factors; they also represent a significanthealthcare burden in the general population and are a major cause ofmissed days at school and lost time in the workplace (Rollinger, J. M.and Schmidtke, M., Med. Res. Rev., 2010, Doi 10.1002/med.20176). Theseconsiderations make it clear that novel medicines, that possess improvedefficacy over current therapies, are urgently required to prevent andtreat rhinovirus-mediated upper respiratory tract infections. In generalthe strategies adopted for the discovery of improved antiviral agentshave targeted various proteins produced by the virus, as the point oftherapeutic intervention. However, the wide range of rhinovirusserotypes makes this a particularly challenging approach to pursue andmay explain why, at the present time, a medicine for the prophylaxis andtreatment of rhinovirus infections has yet to be approved by anyregulatory agency.

Viral entry into the host cell is associated with the activation of anumber of intracellular signalling pathways controlled by the relativeactivation and inactivation of specific kinases which are believed toplay a prominent role in the initiation of inflammatory processes(reviewed by Ludwig, S, 2007; Signal Transduction, 7:81-88) and of viralpropagation and subsequent release.

It has been disclosed previously that compounds that inhibit theactivity of both c-Src and Syk kinases are effective agents againstrhinovirus replication (Charron, C. E. et al., WO 2011/158042) and thatcompounds that inhibit p59-HCK are effective against influenza virusreplication (Charron, C. E. et al., WO 2011/070369). For the reasonssummarized above, in combination with the inhibition of p38 MAPKs, theseare particularly advantageous inherent properties for compounds designedto treat chronic respiratory diseases.

Certain p38 MAPK inhibitors have also been described as inhibitors ofthe replication of respiratory syncytial virus (Cass, L. et al., WO2011/158039).

The precise etiology of IBD is uncertain, but is believed to be governedby genetic and environmental factors that interact to promote anexcessive and poorly controlled mucosal inflammatory response directedagainst components of the luminal microflora. This response is mediatedthrough infiltration of inflammatory neutrophils, dendritic cells andT-cells from the periphery. Due to the ubiquitous expression of p38 ininflammatory cells it has become an obvious target for investigation inIBD models. Studies investigating the efficacy of p38 inhibitors inanimal models of IBD and human biopsies from IBD patients indicated thatp38 could be a target for the treatment of IBD (Hove, T. ten et al.,Gut, 2002, 50:507-512, Docena, G. et al., J. of Trans. Immunol., 2010,162:108-115). However, these findings are not completely consistent withother groups reporting no effect with p38 inhibitors (Malamut G. et al.,Dig. Dis. Sci, 2006, 51:1443-1453). A clinical study in Crohn's patientsusing the p38 alpha inhibitor BIRB796 demonstrated potential clinicalbenefit with an improvement in C-reactive protein levels. However thisimprovement was transient, returning to baseline by week 8 (Schreiber,S. et al., Clin. Gastro. Hepatology, 2006, 4:325-334). A small clinicalstudy investigating the efficacy of CNI-1493, a p38 and Jnk inhibitor,in patients with severe Crohn's disease showed significant improvementin clinical score over 8 weeks (Hommes, D. et al. Gastroenterology. 2002122:7-14).

T cells are known to play key role in mediating inflammation of thegastrointestinal tract. Pioneering work by Powrie and colleaguesdemonstrated that transfer of naive CD4+ cells into severely compromisedimmunodeficient (SCID) animals results in the development of colitiswhich is dependent on the presence of commensal bacteria (Powrie F. etal. Int Immunol. 1993 5:1461-71). Furthermore, investigation of mucosalmembranes from IBD patients showed an upregulation of CD4+ cells whichwere either Th1 (IFNγ/IL-2) or Th2 (IL5/TGFβ) biased depending onwhether the patient had Crohn's disease or ulcerative colitis (Fuss I J.et al. J Immunol. 1996 157:1261-70). Similarly, T cells are known toplay a key role in inflammatory disorders of the eye with severalstudies reporting increased levels of T cell associated cytokines (IL-17and IL-23) in sera of Behçets patients (Chi W. et al. Invest OphthalmolVis Sci. 2008 49:3058-64). In support, Direskeneli and colleaguesdemonstrated that Behçets patients have increased Th17 cells anddecreased Treg cells in their peripheral blood (Direskeneli H. et al. JAllergy Clin Immunol. 2011 128:665-6).

One approach to inhibit T cell activation is to target kinases which areinvolved in activation of the T cell receptor signalling complex. Sykand Src family kinases are known to play a key role in this pathway,where Src family kinases, Fyn and Lck, are the first signallingmolecules to be activated downstream of the T cell receptor (Barber E K.et al. PNAS 1989 86:3277-81). They initiate the tyrosine phosphorylationof the T cell receptor leading to the recruitment of the Syk familykinase, ZAP-70. Animal studies have shown that ZAP-70 knockout resultsin a SCID phenotype (Chan A C. et al. Science. 1994 10;264(5165):1599-601).

A clinical trial in rheumatoid arthritis patients with the Syk inhibitorFostamatinib demonstrated the potential of Syk as an anti-inflammatorytarget with patients showing improved clinical outcome and reduced serumlevels of IL-6 and MMP-3 (Weinblatt M E. et al. Arthritis Rheum. 200858:3309-18). Syk kinase is widely expressed in cells of thehematopoietic system, most notably in B cells and mature T cells.Through interaction with immunoreceptor tyrosine-based activation (ITAM)motifs it plays an important role in regulating T cell and B cellexpansion as well as mediating immune-receptor signalling ininflammatory cells. Syk activation leads to IL-6 and MMPrelease—inflammatory mediators commonly found upregulated ininflammatory disorders including IBD and rheumatoid arthritis (Wang Y D.et al World J Gastroenterol 2007; 13: 5926-5932, Litinsky I et al.Cytokine. 2006 January 33:106-10).

In addition to playing key roles in cell signalling events which controlthe activity of pro-inflammatory pathways, kinase enzymes are now alsorecognised to regulate the activity of a range of cellular functions.Among those which have been discussed recently are the maintenance ofDNA integrity (Shilo, Y. Nature Reviews Cancer, 2003, 3:155-168) andco-ordination of the complex processes of cell division. An illustrationof recent findings is a publication describing the impact of a set ofinhibitors acting upon the so-called “Olaharsky kinases” on thefrequency of micronucleus formation in vitro (Olaharsky, A. J. et al.,PLoS Comput. Biol., 2009, 5(7):e1000446). Micronucleus formation isimplicated in, or associated with, disruption of mitotic processes andis therefore an undesirable manifestation of potential toxicity.Inhibition of glycogen synthase kinase 3α (GSK3α) was found to be aparticularly significant factor that increases the likelihood of akinase inhibitor promoting micronucleus formation. Recently, inhibitionof the kinase GSK3β with RNAi was also reported to promote micronucleusformation (Tighe, A. et al., BMC Cell Biology, 2007, 8:34).

It may be possible to attenuate the adverse effects arising from druginteractions with Olaharsky kinases, such as GSK3α, by optimisation ofthe dose and/or by changing the route of administration. However, itwould be more advantageous to identify therapeutically useful moleculesthat demonstrate low or undetectable activity against these off-targetenzymes and consequently elicit little or no disruption of mitoticprocesses, as measured in mitosis assays.

It is evident from consideration of the literature cited hereinabovethat there remains a need to identify and develop new p38 MAP kinaseinhibitors that have improved therapeutic potential over currentlyavailable treatments. Desirable compounds are those that exhibit asuperior therapeutic index by exerting, at the least, an equallyefficacious effect as previous agents but, in one or more respects, areless toxic at the relevant therapeutic dose. The present inventiontherefore, inter alia, provides such novel compounds that inhibit theenzyme activity of p38 MAP kinase, for example with certain sub-typespecificities, optionally together with Syk kinase and tyrosine kinaseswithin the Src family (particularly c-Src) thereby possessing goodanti-inflammatory properties, and suitable for use in therapy.

In one or more embodiments the compounds exhibit a long duration ofaction and/or persistence of action.

SUMMARY OF THE INVENTION

In a first aspect, the invention provides a compound of formula (I),

wherein:Q represents thienyl, phenyl or pyridinyl, either of which mayoptionally bear 1 to 3 substituents independently selected from,hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₁₋₆hydroxyalkyl, NH₂, N(H)—C₁₋₆ alkyl, N(C₁₋₆ alkyl)₂, -L-P(O)R′R″, C₁₋₆alkylene-5-10 membered heterocycle and C₀₋₃ alkylene-O—C₀₋₆alkylene-5-10 membered heterocycle;L is a direct bond or C₁₋₂ alkylene;R′ represents C₁₋₄ alkyl;R″ represents C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₁₋₄ alkoxy or hydroxy;or R′ and R″ together combine to form C₃₋₆ n-alkylene, wherein one CH₂of said n-alkylene group is optionally replaced by O, N(H) or N(C₁₋₄alkyl);X represents CH or N,Y represents NR²R³;

R is

-   -   C₁₋₆ alkyl,    -   C₂₋₆ alkenyl,    -   C₁₋₆ hydroxyalkyl,    -   C₁₋₆ haloalkyl,    -   C₁₋₆ alkyl substituted by C₂₋₃ alkynyl, C₁₋₃ alkoxy or cyano,    -   C₀₋₂ alkylene-C₃₋₈ cycloalkyl optionally substituted with C₁₋₃        alkyl,    -   a 4-5 membered heterocycle optionally substituted with C₁₋₃        alkyl or    -   Si(R^(1a))(R^(1b))(R^(1c));        R^(1a) and R^(1b) independently represent C₁₋₄ alkyl or C₃₋₆        cycloalkyl, or R^(1a) and R^(1b) together combine to form C₂₋₆        alkylene;        R^(1c) represents C₁₋₂ alkyl;        R^(a) and R^(b), together with the C-atoms to which they are        attached, form a fused phenyl ring that is optionally        substituted by one or more substituents selected from C₁₋₃        alkyl, C₁₋₃ haloalkyl, cyano and halo,        or one of R^(a) and R^(b) represents H, halo, cyano, C₁₋₃ alkyl        or C₁₋₃ haloalkyl and the other independently represents halo,        cyano, C₁₋₃ alkyl or C₁₋₃ haloalkyl        or R^(a) and R^(b) together represent C₃₋₅ n-alkylene, which        alkylene group is optionally substituted by one or more methyl        substituents and/or which alkylene group optionally contains one        C—C double bond between two C-atoms of the n-alkylene chain;        R¹ is selected from hydrogen, OH, halogen, CN, C₁₋₆ alkyl, C₂₋₆        alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₀₋₃ alkylene-C₃₋₆        cycloalkyl, C₀₋₃ alkylene-O—C₁₋₃ alkylene-C₃₋₆ cycloalkyl, C₁₋₆        alkoxy, C₁₋₆ haloalkoxy, C₁₋₆ hydroxyalkyl, C₀₋₃        alkylene-SO₂C₁₋₃alkyl, C₀₋₃ alkylene-SO₂NR⁴R⁵, and C₀₋₃        alkylene-NR⁶R⁷ and C₀₋₃ alkylene-NCOR⁶R⁷;        one of R² and R³ represents —[C₂₋₄ alkylene-O]₁₋₁₂—[C₂₋₄        alkylene]-R^(2a) and the other of R² and R³ is selected from H,        C₁₋₈ alkyl, C₀₋₆ alkylene aryl, C₀₋₆ alkylene heteroaryl, —[C₂₋₄        alkylene-O]₀₋₁₂—[C₂₋₄ alkylene]-R^(2a), C₀₋₆ alkylene-4-10        membered heterocycle, and C₀₋₃ alkylene-O—C₀₋₆ alkylene-4-10        membered heterocycle with the proviso that when the said        heterocycle is linked through nitrogen there are at least two        C-atoms in the alkylene chain that links that nitrogen atom to        the essential O atom of the substituent, wherein independently        each alkyl or alkylene group optionally bears 1 oxo substituent,        and optionally one or two carbon atoms in the alkyl or alkylene        chain may each be replaced by a heteroatom selected from O, N or        S(O)_(p), such that when said alkyl or alkylene comprises an        amine said amino group is a tertiary amine,        wherein each 4-10 membered heterocycle is optionally substituted        by 1 or 2 groups independently selected from halo, OH, C₁₋₆        alkyl, C₁₋₄ haloalkyl, C₀₋₃ alkylene-O—C₀₋₆ alkyl, C₀₋₃        alkylene-O—C₁₋₃ haloalkyl, C₀₋₆ alkylene aryl, C₀₋₃        alkylene-O—C₀₋₃ alkylene aryl, C₀₋₆ alkylene heteroaryl, C₀₋₃        alkylene-O—C₀₋₃ alkylene heteroaryl, C(O)C₁₋₆ alkyl, SO₂NR⁸R⁹,        and C₀₋₃ alkylene-NR⁸R⁹, C₀₋₃ alkylene-NR⁸SO₂R⁹ and C₀₋₃        alkylene-NR⁸C(O)R⁹;        R^(2a) represents OR^(2b) or N(R^(2c))R^(2d);        R^(2b) to R^(2d) independently represent H or C₁₋₄ alkyl        optionally substituted by one or more halo atoms, or R^(2c) and        R^(2d) together represent    -   C₃₋₆ n-alkylene,    -   C₄₋₆ n-alkylene interrupted between C2 and C3 by —O— or        —N(R^(2e))— or    -   C₆ n-alkylene interrupted between C2 and C3, or between C3 and        C4, by —O— or —N(R^(2e))—,        any of which n-alkylene groups are optionally substituted by one        or more substituents selected from halo, hydroxy, oxo, C₁₋₄        alkyl and C₁₋₄ alkoxy;        R^(2e) represents H or C₁₋₆ alkyl optionally substituted by one        or more substituents selected from halo and hydroxy;        R⁴ is H or C₁₋₄ alkyl;        R⁵ is H or C₁₋₄ alkyl,        R⁶ is H or C₁₋₄ alkyl, C(O)C₁₋₃alkyl and SO₂C₁₋₃ alkyl;        R⁷ is H or C₁₋₄ alkyl, C(O)C₁₋₃alkyl and SO₂C₁₋₃ alkyl;        R⁸ is H or C₁₋₄ alkyl, and        R⁹ is H or C₁₋₄ alkyl,        p is 0, 1 or 2        or a pharmaceutically acceptable salt thereof, including all        stereoisomers and tautomers thereof.

Compounds of the invention are inhibitors of p38 MAP kinase especiallyof the alpha sub-type.

In at least some embodiments compounds of the present invention have lowB-Raf binding, for example less than 40% inhibition of the kinasebinding at 500 nM, such as 30% inhibition or less in an assay such asthe Kinomescan method.

B-Raf is a member of the Raf kinase family of serine/threonine-specificprotein kinases. This protein plays a role in regulating the MAPkinase/ERKs signalling pathway, which affects cell division,differentiation, and secretion. A mutation of the gene has beenassociated with cancer in humans (Davies, H. et al., Nature, 2002,417(6892):949-54).

Cell signalling can bypass selective inhibition of B-Raf withundesirable consequences (Lo, R. S., Cell Research, advance onlinepublication 8 May 2012; doi: 10.1038/cr.2012.78). It is thereforepreferable that kinase inhibitors intended for use as anti-inflammatorymedicines should have minimal potential to interact with B-Raf.

The present compounds also display low affinity for GSK3α kinase inbinding assays, which is considered to be beneficial in a therapeuticcontext, in particular in relation to minimising toxicity in vivo.

In at least some embodiments, compounds of the present invention havep59-HCK inhibitory activity which may also augment their advantageoustherapeutic profile.

DETAILED DESCRIPTION OF THE INVENTION

Alkyl as used herein refers to straight chain or branched chain alkyl,such as, without limitation, methyl, ethyl, n-propyl, iso-propyl, butyl,n-butyl and tert-butyl. In one embodiment alkyl refers to straight chainalkyl.

Alkoxy as used herein refers to straight or branched chain alkoxy, forexample methoxy, ethoxy, propoxy, butoxy. Alkoxy as employed herein alsoextends to embodiments in which the or an oxygen atom (e.g. a singleoxygen atom) is located within the alkyl chain, for example —C₁₋₃alkylOC₁₋₃ alkyl, such as —CH₂CH₂OCH₃ or —CH₂OCH₃. Thus in oneembodiment the alkoxy is linked through carbon to the remainder of themolecule, for example —C_(6-n)alkyl-O—C_(6-m)alkyl in which n=1-5, m=1-5and n+m=6-10. In one embodiment the alkoxy is linked through oxygen tothe remainder of the molecule, for example —OC₁₋₆ alkyl. In oneembodiment the disclosure relates to straight chain alkoxy. In oneembodiment the alkoxy is linked through oxygen to the remainder of themolecule but the alkoxy group contains a further oxygen atom, forexample —OCH₂CH₂OCH₃.

Halo or halogen includes fluoro, chloro, bromo or iodo, in particularfluoro, chloro or bromo, especially fluoro or chloro.

Alkyl substituted by halo (haloalkyl) as employed herein refers to alkylgroups having 1 to 6 halogen atoms, for example 1 to 5 halogens, such asper haloalkyl, in particular perfluoroalkyl, more specifically —CF₂CF₃or CF₃.

Alkyl substituted by hydroxy (hydroxyalkyl) as employed herein refers toalkyl groups having 1 to 3 hydroxy groups, for example 1 or 2 hydroxysubstituents thereon, for example —CH₂CH₂OH, —C(CH₃)CH₂OH, —C(CH₃)₂CH₂OHor similar.

Alkoxy substituted by halo (haloalkoxy) as employed herein refers toalkoxy groups having 1 to 6 halogen atoms, for example 1 to 5 halogens,such as per haloalkoxy, in particular perfluoroalkoxy, more specifically—OCF₂CF₃ or —OCF₃.

Unless otherwise specified, alkylene as employed herein is a straightchain or branched chain carbon linking group, for example comprisingmethylenes, between two other moieties. It will be clear to thoseskilled in the art that groups defined as, for example C₂₋₈ alkenyl andC₂₋₈ alkynyl may comprise an alkylene portion. For the avoidance ofdoubt, the term “n-alkylene”, when used herein, refers to straight chainalkylene.

It will be clear to persons skilled in the art that the heteroatom mayreplace a primary, secondary or tertiary carbon, that is a CH₃, —CH₂— ora —CH—, group, as technically appropriate and hydrogen or branching inthe alkyl or alkylene chain will fill the valency of the heteroatom asappropriate to the location, for example where a terminal primary carbonis replaced by an oxygen heteroatom the terminal group will be analcohol.

C₁₋₆ alkyl includes C₁, C₂, C₃, C₄, C₅ and C₆.

C₁₋₆ alkoxy includes C₁, C₂, C₃, C₄, C₅ and C₆.

The term 5-10 membered heterocycle, as employed herein refers to a 5 to10 membered saturated or partially unsaturated non-aromatic ringcomprising one or more, for example 1, 2, 3 or 4 heteroatomsindependently selected from O, N and S, wherein optionally one or twocarbons in the ring may bear an oxo substituent. Any valencies of aheteroatom not employed in forming or retaining the ring structure maybe filled by hydrogen or a substituent, as appropriate. Thus theoptional substituents on the heterocycles may be attached to a carbon oron a heteroatom, such as nitrogen as appropriate. Examples of 5-10membered heterocycles include, pyrroline, pyrrolidine, tetrahydrofuran,thiepane, oxepane piperidine, piperazine, morpholine, thiomorpholine,dioxane, tetrahydrothiophene, pyrazoline, imidazoline, pyrazolidine,oxoimidazolidine, dioxolane, thiazolidine, isoxazolidine, dihydropyran,dihydroindene, dihydroisobenzofuran, isoindolin-1-one, chroman,1,2,3,4-tetrahydroquinoline, 2,3-dihydrobenzo[b][1,4]dioxineazocane, andthe like.

The term 5-6 membered heterocycle as employed herein refers to a 5 to 6membered saturated or partially unsaturated non-aromatic ring comprisingone or more, for example 1, 2, 3 or 4 heteroatoms independently selectedfrom O, N and S wherein optionally one or two carbons in the ring maybear an oxo substituent. The definition of C₅₋₆ heterocycle as employedherein refers to a 5 to 6 membered saturated or partially unsaturatednon-aromatic carbocyclic ring comprising one or more, for example 1, 2,3 or 4 heteroatoms independently selected from O, N and S, wherein eachheteroatom replaces a carbon atom and optionally one or two carbons maybear an oxo substituent. Clearly any valencies of a heteroatom notemployed in forming or retaining the ring structure may be filled byhydrogen or a substituent, as appropriate. Thus substituents onheterocycles may be on carbon or on a heteroatom, such as N asappropriate. Examples of heterocycles and C₅₋₆ heterocycles includepyrroline, pyrrolidine, tetrahydrofuran, tetrahydrothiophene,pyrazoline, imidazoline, pyrazolidine, imidazolidine, oxoimidazolidine,dioxolane, thiazolidine, isoxazolidine, pyran, dihydropyran, piperidine,piperazine, morpholine, dioxane, thiomorpholine and oxathiane.

When employed herein, the group morpholinyl suitably representsN-morpholinyl.

In one embodiment there is provided a compound of formula (Ia1) or,particularly, formula (Ia2):

wherein R, R^(a), R^(b), R¹, Q and Y are defined as above for compoundsof formula (I).

In one embodiment there is provided a compound of formula (Ib1) or,particularly, formula (Ib2):

wherein R, R^(a), R^(b), R¹, Q, X and Y are defined above for compoundsof formula (I)

In one embodiment there is provided a compound of formula (Ic1) or,particularly, formula (Ic2):

wherein R, R^(a), R^(b), R¹, Q and Y are defined above for compounds offormula (I).

In one embodiment there is provided a compound of formula (Id1) or,particularly, formula (Id2):

wherein R, R^(a), R^(b), R¹, Q and Y are defined above for compounds offormula (I).

In one embodiment there is provided a compound of formula (Ie1) or,particularly, formula (Ie2):

wherein R, R^(a), R^(b), R¹, Q and Y are defined above for compounds offormula (I).

In one embodiment there is provided a compound of formula (If1) or,particularly, formula (If2):

wherein R, R^(a), R^(b), R¹, Q and Y are defined above for compounds offormula (I).

In one embodiment there is provided a compound of formula (Ig1) or,particularly, formula (Ig2):

wherein R, R^(a), R^(b), R¹, X, Q and Y are as defined above forcompounds of formula (I).

In one embodiment there is provided a compound of formula (Ih1) or,particularly, formula (Ih2):

wherein R, R^(a), R^(b), R¹, X, Q and Y are as defined above forcompounds of formula (I).

Generally in substituents such C₀₋₃ alkylene-O—C₀₋₆ alkylene-5-10membered heterocycle, for example as defined for R² or R³, when the saidheterocycle is linked through nitrogen the group will then be defined asC₀₋₃ alkylene-O—C₂₋₆ alkylene-5-10 membered heterocycle.

Generally when Q comprises a phenyl or pyridine substituted with a C₁₋₆alkylene-5-10 membered heterocycle or C₀₋₃ alkylene-O—C₀₋₆ alkylene-5-10membered heterocycle then R² and R³ are independently selected from H,C₁₋₈ alkyl, wherein independently each alkyl or alkylene groupoptionally bears 1 oxo substituent, and optionally up to two carbonatoms in the alkyl or alkylene chain may be replaced by a heteroatomselected from O, N or S(O)_(p), such that when alkyl or alkylenecomprises an amine said amino group is a tertiary amine.

In one embodiment Q represents phenyl bearing one or two substituentsindependently selected from hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy,C₁₋₆ haloalkoxy, C₁₋₆ hydroxyalkyl, N(C₁₋₆ alkyl)₂, C₁₋₆ alkylene-5-10membered heterocycle and C₀₋₃ alkylene-O—C₁₋₆ alkylene-5-10 memberedheterocycle (e.g. one or two substituents independently selected fromhydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₁₋₆hydroxyalkyl, C₁₋₆ alkylene-5-10 membered heterocycle and C₀₋₃alkylene-O—C₁₋₆ alkylene-5-10 membered heterocycle).

In one embodiment Q represents phenyl bearing a methyl, methoxy,—N(CH₃)₂ or —OCH₂CH₂OCH₃ (e.g. methyl, methoxy, or —OCH₂CH₂OCH₃), forexample one of said substituents, in particular in the para position.

In one embodiment Q is dimethyl phenyl, for example where the methylsubstituents are in the meta and para position.

In one embodiment Q represents pyridinyl bearing one substituentindependently selected from hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy,C₁₋₆ haloalkoxy, C₁₋₆ hydroxyalkyl, C₁₋₆ alkylene-5-10 memberedheterocycle and C₀₋₃ alkylene-O—C₁₋₆ alkylene-5-10 membered heterocycle.

In one embodiment Q is methoxypyridinyl, for example6-methoxypyridin-3-yl.

In one embodiment Q represents thienyl optionally bearing onesubstituent independently selected from hydroxyl, halogen, C₁₋₆ alkyl,C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₁₋₆ hydroxyalkyl, C₁₋₆ alkylene-5-10membered heterocycle and C₀₋₃ alkylene-O—C₁₋₆ alkylene-5-10 memberedheterocycle.

In one embodiment R is ethyl, isopropyl, tert-butyl, cyclopropyl,1-methylcyclopropyl, propen-2-yl, CF₃, C₂F₅, oxetanyl, (methyl)oxetanylor tetrahydrofuranyl (e.g. ethyl, isopropyl, tert-butyl, cyclopropyl,1-methylcyclopropyl, CF₃, C₂F₅, oxetanyl, (methyl)oxetanyl ortetrahydrofuranyl), such as isopropyl or tert-butyl.

In one embodiment R is C(CH₃)₂CH₂OH or CH(CH₃)CH₂OH.

In one embodiment R is 1-hydroxy-2-methylpropan-2-yl.

In one embodiment R¹ is H, Br, Cl, CH₃, CH₂CH₃, CN, N(CH₃)₂, CF₃,ethynyl, OCH₃, OCHF₂, OCH₂CH₃ or OCH₂(CH₃)₂ (e.g. R¹ is H, Br, Cl, CH₃,CN, N(CH₃)₂, CF₃, ethynyl, OCH₃, OCH₂CH₃ or OCH₂(CH₃)₂).

In one embodiment R⁴ is H or methyl.

In one embodiment R⁵ is H or methyl.

In one embodiment R⁶ is H or methyl.

In one embodiment R⁷ is H or methyl.

In one embodiment R⁸ is H or methyl.

In one embodiment R⁹ is H or methyl.

Embodiments of the invention that may be mentioned include compounds offormulae (I), (Ia1), (Ia2), (Ib1), (Ib2), (Ic1), (Ic2), (Id1), (Id2),(Ie1), (Ie2), (If1), (If2), (Ig1), (Ig2), (Ih1) and (Ih2) wherein:

Q represents phenyl substituted by -L-P(O)R′R″ or, particularly, phenylbearing one or two substituents independently selected from hydroxyl,halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₁₋₆ hydroxyalkyl,N(C₁₋₆ alkyl)₂, C₁₋₆ alkylene-5-10 membered heterocycle and C₀₋₃alkylene-O—C₁₋₆ alkylene-5-10 membered heterocycle (e.g. Q representsphenyl mono-substituted (e.g. in the para position) by methyl, methoxy,—N(CH₃)₂ or —OCH₂CH₂OCH₃ or di-substituted (e.g. in the meta and parapositions) by methyl),

or Q represents pyridinyl bearing one substituent independently selectedfrom hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₁₋₆hydroxyalkyl, C₁₋₆ alkylene-5-10 membered heterocycle and C₀₋₃alkylene-O—C₁₋₆ alkylene-5-10 membered heterocycle (e.g. Q representsmethoxypyridinyl, such as 6-methoxypyridin-3-yl);

L is CH₂;

R′ and R″ both represent C₁₋₄ alkyl (e.g. C₁₋₂ alkyl, such as methyl);R^(a) and R^(b) together represent —(CH₂)₃₋₅- or, particularly, R^(a)and R^(b), together with the C-atoms to which they are attached, form afused phenyl ring, or one of R^(a) and R^(b) represents halo,C₁₋₃ alkyl or C₁₋₃ haloalkyl and the other independently representshalo, cyano, C₁₋₃ alkyl orC₁₋₃ haloalkyl (e.g. R^(a) and R^(b) both represent methyl, fluoro orchloro);Y represents NR²R³;R² represents —[C₂₋₄ alkylene-O]₁₋₆—[C₂₋₄ alkylene]-R^(2a) (e.g. —[C₂₋₄alkylene-O]₁₋₃—[C₂₋₄ alkylene]-R^(2a), such as —[CH₂CH₂O]₁₋₂—CH₂CH₂R^(2a));R³ represents methyl or, particularly, H;R^(2a) represents OR^(2b) or N(R^(2c))R^(2d);R^(2b) represents H or C₁₋₄ alkyl (e.g. R^(2b) represents methyl);R² and R^(2d) independently represent H or C₁₋₄ alkyl (e.g. methyl) orR^(2c) and R^(2d) together represent C₄₋₅ n-alkylene, which n-alkylenegroup is optionally interrupted between C2 and C3 by —O— or —N(R^(2e))—(e.g. R^(2c) and R^(2d) either both represent methyl or togetherrepresent —CH₂CH₂—O—CH₂CH₂—, —CH₂CH₂—NH—CH₂CH₂— or—CH₂CH₂—N(CH₃)—CH₂CH₂—);R^(2e) represents H or C₁₋₄ alkyl (e.g. methyl);R represents

-   -   Si(C₁₋₂ alkyl)₃ (e.g. Si(CH₃)₃),    -   —C(C₁₋₂ alkyl)₂-C₂₋₃ alkynyl (e.g. —C(CH₃)₂—C≡C—H) or,        particularly,    -   C₁₋₆ alkyl optionally substituted by hydroxy, cyano or methoxy        or by one or more fluoro groups,    -   C₂₋₆ alkenyl or    -   C₃₋₄ cycloalkyl, which latter group is optionally substituted by        C₁₋₃ alkyl        (e.g. R represents ethyl, isopropyl, n-propyl, tert-butyl,        cyclopropyl, 1-methylcyclopropyl, CF₃, C₂F₅, —C(CH₃)₂CF₃        oxetanyl, (methyl)oxetanyl, tetrahydrofuranyl or propen-2-yl,        such as isopropyl, propen-2-yl or tert-butyl); and/or        R¹ represents H, halogen (e.g. F, Br or Cl), CN, C₁₋₄ alkyl        (e.g. methyl or ethyl), C₂₋₄ alkynyl (e.g. ethynyl), C₁₋₄        fluoroalkyl (e.g. CF₃), C₁₋₄ alkoxy (e.g. OCH₃, OCH₂CH₃ or        OCH₂(CH₃)₂), C₁₋₄ haloalkoxy (e.g. OCHF₂) or NR⁶R⁷ (e.g.        N(CH₃)₂) (e.g. R¹ represents H, halogen (e.g. F, Br or Cl), CN,        C₁₋₄ alkyl (e.g. methyl or ethyl), C₂₋₄ alkynyl (e.g. ethynyl),        C₁₋₄ fluoroalkyl (e.g. CF₃), C₁₋₄ alkoxy (e.g. OCH₃, OCH₂CH₃ or        OCH₂(CH₃)₂), or NR⁶R⁷ (e.g. N(CH₃)₂), in particular R¹        represents ethynyl or OCH₃).

More particular embodiments of the invention that may be mentionedinclude compounds of formulae (I), (Ia1), (Ia2), (Ib1), (Ib2), (Ic1),(Ic2), (Id1), (Id2), (Ie1), (Ie2), (If1), (If2), (Ig1), (Ig2), (Ih1) and(Ih2) wherein:

Q represents phenyl mono-substituted (e.g. in the meta position) by—CH₂—P(O)(C₁₋₂ alkyl)₂ or, particularly, phenyl mono-substituted (e.g.in the para position) by C₁₋₆ alkyl (e.g. methyl), C₁₋₆ alkoxy (e.g.methoxy), C₁₋₆ haloalkoxy or N(C₁₋₆ alkyl)₂ (e.g. N(CH₃)₂) (for example,Q represents phenyl substituted in the para position by methyl, methoxyor dimethylamino);R^(a) and R^(b) together represent —(CH₂)₄— or, particularly, R^(a) andR^(b), together with the C-atoms to which they are attached, form afused phenyl ring;Y represents NR²R³;R² represents —[C₂₋₃ alkylene-O]₁₋₃—[C₂₋₃ alkylene]-R^(2a) (e.g. R²represents —[CH₂CH₂O]₁₋₂—

CH₂CH₂R^(2a));

R³ represents H;R^(2a) represents —O—(C₁₋₃ alkyl) (e.g. —OCH₃) or N(R^(2c))R^(2d);R^(2c) and R^(2d) independently represent H or C₁₋₃ alkyl (e.g. methyl)or R^(2c) and R^(2d) together represent C₄ n-alkylene, which n-alkylenegroup is optionally interrupted between C2 and C3 by —O— or —N(R^(2e))—(e.g. R^(2c) and R^(2d) either both represent methyl or togetherrepresent —CH₂CH₂—N(CH₃)—CH₂CH₂— or, particularly, —CH₂CH₂—O—CH₂CH₂—);R represents C₁₋₄ alkyl optionally substituted by one or more fluorogroups, C₃₋₄ alkenyl or C₃₋₄ cycloalkyl, which latter group isoptionally substituted by methyl (e.g. R represents ethyl, cyclopropyl,CF₃, C₂F₅, —C(CH₃)₂CF₃ or, particularly, isopropyl, 1-methylcyclopropyl,propen-2-yl or tert-butyl); and/orR¹ represents Br, Cl, CN, methyl, ethyl, CF₃, OCHF₂, OCH₂CH₃,OCH₂(CH₃)₂, N(CH₃)₂ or, particularly, ethynyl or OCH₃ (e.g. R¹represents Br, Cl, CN, methyl, ethyl, CF₃, OCH₂CH₃, OCH₂(CH₃)₂, N(CH₃)₂or, particularly, ethynyl or OCH₃).

Particular embodiments of the invention include the following.

-   (1) A compound of formula (I), (Ia1), (Ia2), (Ib1), (Ib2), (Ic1),    (Ic2), (Id1), (Id2), (Ie1), (Ie2), (If1), (If2), (Ig1), (Ig2), (Ih1)    or (Ih2) as defined above, or a pharmaceutically acceptable salt    thereof.-   (2) A compound or salt according to Embodiment (1), wherein Q    represents phenyl bearing one or two substituents independently    selected from hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆    haloalkoxy, C₁₋₆ hydroxyalkyl, N(C₁₋₆ alkyl)₂, C₁₋₆ alkylene-5-10    membered heterocycle and C₀₋₃ alkylene-O—C₁₋₆ alkylene-5-10 membered    heterocycle (e.g. one or two substituents independently selected    from hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,    C₁₋₆ hydroxyalkyl, C₁₋₆ alkylene-5-10 membered heterocycle and C₀₋₃    alkylene-O—C₁₋₆ alkylene-5-10 membered heterocycle).-   (3) A compound or salt according to Embodiment (1) or Embodiment    (2), wherein Q represents phenyl bearing a methyl, methoxy, —N(CH₃)₂    or —OCH₂CH₂OCH₃ (e.g. a methyl, methoxy or —OCH₂CH₂OCH₃).-   (4) A compound or salt according to any one of Embodiments (1) to    (3), wherein Q represents phenyl substituted in the para position by    methyl, methoxy, —N(CH₃)₂ or —OCH₂CH₂OCH₃ (e.g. by methyl, methoxy    or —OCH₂CH₂OCH₃).-   (5) A compound or salt according to any one of Embodiments (1) to    (3), wherein Q is dimethyl phenyl, for example where the methyl    substituents are in the meta and para position.-   (7) A compound or salt according to Embodiment (1), wherein Q    represents pyridinyl bearing one substituent independently selected    from hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,    C₁₋₆ hydroxyalkyl, C₁₋₆ alkylene-5-10 membered heterocycle and C₀₋₃    alkylene-O—C₁₋₆ alkylene-5-10 membered heterocycle.-   (8) A compound or salt according to Embodiment (7), wherein Q is    methoxypyridinyl, for example 6-methoxypyridin-3-yl.-   (9) A compound or salt according to Embodiment (1), wherein Q    represents thienyl optionally bearing one substituent independently    selected from hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆    haloalkoxy, C₁₋₆ hydroxyalkyl, C₁₋₆ alkylene-5-10 membered    heterocycle and C₀₋₃ alkylene-O—C₁₋₆ alkylene-5-10 membered    heterocycle.-   (10) A compound or salt according to any one of Embodiments (1) to    (9), wherein R is ethyl, isopropyl, tert-butyl, cyclopropyl,    1-methylcyclopropyl, CF₃, C₂F₅, oxetanyl, (methyl)oxetanyl or    tetrahydrofuranyl, such as isopropyl or tert-butyl.-   (11) A compound or salt according to any one of Embodiments (1) to    (9), wherein R is C(CH₃)₂CH₂OH or CH(CH₃)CH₂OH.-   (12) A compound or salt according to any one of Embodiments (1) to    (9), wherein R is 1-hydroxy-2-methylpropan-2-yl.-   (13) A compound or salt according to any one of Embodiments (1) to    (12), wherein R¹ is H, Br, Cl, CH₃, CH₂CH₃, CN, N(CH₃)₂, CF₃,    ethynyl, OCH₃, OCHF₂, OCH₂CH₃ or OCH₂(CH₃)₂ (e.g. R¹ is H, Br, Cl,    CH₃, CN, N(CH₃)₂, CF₃, ethynyl, OCH₃, OCH₂CH₃ or OCH₂(CH₃)₂).-   (14) A compound or salt according to any one of Embodiments (1) to    (13), wherein R⁴ is H or methyl.-   (15) A compound or salt according to any one of Embodiments (1) to    (14), wherein R⁵ is H or methyl.-   (16) A compound or salt according to any one of Embodiments (1) to    (15), wherein R⁶ is H or methyl.-   (17) A compound or salt according to any one of Embodiments (1) to    (16), wherein R⁷ is H or methyl.-   (18) A compound or salt according to any one of Embodiments (1) to    (17), wherein R⁸ is H or methyl.-   (19) A compound or salt according to any one of Embodiments (1) to    (18), wherein R⁹ is H or methyl.-   (20) A compound or salt according to any one of Embodiments (1)    and (10) to (19), wherein Q represents phenyl mono-substituted (e.g.    in the para position) by C₁₋₆ alkyl (e.g. methyl), C₁₋₆ alkoxy (e.g.    methoxy), C₁₋₆ haloalkoxy or N(C₁₋₆ alkyl)₂ (e.g. N(CH₃)₂).-   (21) A compound or salt according to Embodiment (20), wherein Q    represents phenyl substituted in the para position by methyl,    methoxy or dimethylamino.-   (22) A compound or salt according to any one of Embodiments (1) to    (21), wherein R^(a) and R^(b), together with the C-atoms to which    they are attached, form a fused phenyl ring.-   (23) A compound or salt according to any one of Embodiments (1)    to (22) (e.g. any one of Embodiments (1) to (21)), wherein R²    represents —[C₂₋₃ alkylene-O]₁₋₃—[C₂₋₃ alkylene]-R^(2a) (e.g. R²    represents —[CH₂CH₂O]₁₋₂—CH₂CH₂R^(2a)).-   (24) A compound or salt according to any one of Embodiments (1) to    (23), wherein R³ represents H.-   (25) A compound or salt according to any one of Embodiments (1) to    (24), wherein R^(2a) represents —O—(C₁₋₃ alkyl) (e.g. —OCH₃) or    N(R^(2c))R^(2d).-   (26) A compound or salt according to any one of Embodiments (1) to    (25), wherein R² and R^(2d) independently represent H or C₁₋₃ alkyl    (e.g. methyl) or R^(2c) and R^(2d) together represent C₄ n-alkylene,    which n-alkylene group is optionally interrupted between C2 and C3    by —O— or —N(R^(2e))— (e.g. R^(2c) and R^(2d) either both represent    methyl or together represent —CH₂CH₂—N(CH₃)—CH₂CH₂— or,    particularly, —CH₂CH₂—O—CH₂CH₂—).-   (27) A compound or salt according to any one of Embodiments (1)    to (9) and (13) to (26), wherein R represents C₁₋₄ alkyl optionally    substituted by one or more fluoro groups, C₃₋₄ alkenyl or C₃₋₄    cycloalkyl, which latter group is optionally substituted by methyl    (e.g. R represents ethyl, cyclopropyl, CF₃, C₂F₅, —C(CH₃)₂CF₃ or,    particularly, isopropyl, 1-methylcyclopropyl, propen-2-yl or    tert-butyl).-   (28) A compound or salt according to any one of Embodiments (1)    to (12) and (14) to (27), wherein R¹ represents Br, Cl, CN, methyl,    ethyl, CF₃, OCHF₂, OCH₂CH₃, OCH₂(CH₃)₂, N(CH₃)₂, ethynyl or OCH₃    (e.g. R¹ represents Br, Cl, CN, methyl, ethyl, CF₃, OCH₂CH₃,    OCH₂(CH₃)₂, N(CH₃)₂, ethynyl or OCH₃).-   (29) A compound or salt according to Embodiment (28), wherein R¹    represents ethynyl or OCH₃.-   (30) A compound or salt according to any one of Embodiments (1)    to (29) above, wherein:    -   Q represents thienyl, phenyl or pyridinyl, either of which is        substituted by NH₂, N(H)—C₁₋₆ alkyl, N(C₁₋₆ alkyl)₂ or,        particularly, -L-P(O)R′R″ and is optionally further substituted        by 1 or 2 substituents independently selected from, hydroxyl,        halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₁₋₆        hydroxyalkyl, NH₂, N(H)—C₁₋₆ alkyl, N(C₁₋₆ alkyl)₂, -L-P(O)R′R″,        C₁₋₆ alkylene-5-10 membered heterocycle and C₀₋₃ alkylene-O—C₀₋₆        alkylene-5-10 membered heterocycle;    -   R represents C₂₋₆ alkenyl (e.g. C₃₋₄ alkenyl, such as        propen-2-yl), C₁₋₆ alkyl substituted by C₁₋₃ alkoxy or cyano        (e.g. secondary C₃₋₆ alkyl substituted by methoxy or cyano, such        as —C(CH₃)₂OCH₃ or —C(CH₃)₂CN) or, particularly, C₁₋₆ alkyl        substituted by C₂₋₃ alkynyl (e.g. —C(C₁₋₂ alkyl)₂-C₂₋₃ alkynyl,        such as —C(CH₃)₂—C≡C—H) or Si(R^(1a))(R^(1b))(R^(1c))(e.g.        Si(C₁₋₂ alkyl)₃, such as Si(CH₃)₃); and/or    -   R^(a) and R^(b), together with the C-atoms to which they are        attached, form a fused phenyl ring that is substituted by one or        more substituents selected from C₁₋₃ alkyl, C₁₋₃ haloalkyl,        cyano and halo,    -   or one of R^(a) and R^(b) represents H, halo, cyano, C₁₋₃ alkyl        or C₁₋₃ haloalkyl and the other independently represents halo,        cyano, C₁₋₃ alkyl or C₁₋₃ haloalkyl, or, particularly, R^(a) and        R^(b) together represent C₃₋₅ n-alkylene, which alkylene group        is optionally substituted by one or more methyl substituents        and/or which alkylene group optionally contains one C—C double        bond between two C-atoms of the n-alkylene chain (e.g. R^(a) and        R^(b) together represent C₃₋₄ n-alkylene, such as —(CH₂)₄—).-   (31) A compound or salt according to any one of Embodiments (1)    to (30) above, wherein:    -   Q represents phenyl substituted by -L-P(O)R′R″ (e.g. in the        para- or, particularly, the meta-position relative to the point        of attachment of the phenyl group to the pyrazole group);    -   L is CH₂ or a direct bond;    -   R′ represents C₁₋₂ alkyl (e.g. methyl);    -   R″ represents C₁₋₂ alkyl (e.g. methyl);    -   or R′ and R″ together combine to form C₄₋₅ n-alkylene;    -   R is —C(C₁₋₂ alkyl)₂-C₂₋₃ alkynyl (e.g. —C(CH₃)₂—C≡C—H) or        Si(C₁₋₂ alkyl)₃ (e.g. Si(CH₃)₃); and/or    -   R^(a) and R^(b) together represent C₃₋₄ n-alkylene (e.g.        —(CH₂)₄—).-   (32) A compound or salt according to any one of Embodiments (1)    to (29) above, wherein:    -   Q represents thienyl, phenyl or pyridinyl, either of which may        optionally bear 1 to 3 substituents independently selected from,        hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,        C₁₋₆ hydroxyalkyl, NH₂, N(H)—C₁₋₆ alkyl, N(C₁₋₆ alkyl)₂, C₁₋₆        alkylene-5-10 membered heterocycle and C₀₋₃ alkylene-O—C₀₋₆        alkylene-5-10 membered heterocycle;    -   R is C₁₋₆ alkyl, C₂₋₆ alkenyl, C₁₋₆ hydroxyalkyl, C₁₋₆        haloalkyl, C₁₋₆ alkyl substituted by C₁₋₃ alkoxy or cyano, C₀₋₂        alkylene-C₃₋₈ cycloalkyl optionally substituted with C₁₋₃ alkyl        or a 4-5 membered heterocycle optionally substituted with C₁₋₃        alkyl; and    -   R^(a) and R^(b), together with the C-atoms to which they are        attached, form a fused phenyl ring that is optionally        substituted by one or more substituents selected from C₁₋₃        alkyl, C₁₋₃ haloalkyl, cyano and halo, or one of R^(a) and R^(b)        represents H, halo, cyano, C₁₋₃ alkyl or C₁₋₃ haloalkyl and the        other independently represents halo, cyano, C₁₋₃ alkyl or C₁₋₃        haloalkyl.-   (33) A compound or salt according to any one of Embodiments (1)    to (32) above, wherein one of R² and R³ represents —[C₂₋₃    alkylene-O]₁₋₃—[C₂₋₃ alkylene]-R^(2a), such as —(CH₂CH₂O)₂₋₃CH₃) and    the other of R² and R³ is as defined above in any of Embodiments (1)    to (32) (e.g. the other of R² and R³ is H).-   (34) A compound or salt according to any one of Embodiments (1)    to (33) above, wherein    -   R represents:    -   C₁₋₆ n-alkyl,    -   C₃₋₆ branched alkyl (e.g. C₄₋₆ branched alkyl),    -   C₂₋₆ alkenyl,    -   C₁₋₆ hydroxyalkyl,    -   C₁₋₆ haloalkyl,    -   C₁₋₆ alkyl substituted by C₁₋₃ alkoxy or cyano,    -   C₀₋₂ alkylene-C₃₋₈ cycloalkyl optionally substituted with C₁₋₃        alkyl, or    -   a 4-5 membered heterocycle optionally substituted with C₁₋₃        alkyl    -   (e.g. R represents ethyl, cyclopropyl, CF₃, C₂F₅, —C(CH₃)₂CF₃        or, particularly, 1-methylcyclopropyl, propen-2-yl or        tert-butyl).-   (35) A compound or salt according to any one of Embodiments (1)    to (32) above, wherein one of R² and R³ represents —[C₂₋₃    alkylene-O]₁₋₅—[C₂₋₃ alkylene]-R^(2a) (e.g. —[C₂₋₃    alkylene-O]₁₋₄—[C₂₋₃ alkylene]-R^(2a), such as —(CH₂CH₂O)₂₋₄CH₃) and    the other of R² and R³ is as defined above in any of Embodiments (1)    to (32) (e.g. the other of R² and R³ is H).-   (36) A compound or salt according to Embodiment (35) above, wherein    R¹ represents ethynyl or OCH₃.-   (37) A compound or salt according to Embodiment (35) or    Embodiment (36) above, wherein R² represents —(CH₂CH₂O)₂₋₄CH₃ and R³    is H.-   (38) A compound or salt according to any one of Embodiments (1)    to (37) above, wherein    -   Q represents phenyl substituted in the para position by methyl,        methoxy or dimethylamino and R represents isopropyl or,        particularly, tert-butyl.

Exemplary compounds of formula (I) are selected from the groupconsisting of:

-   3-ethynyl-5-((4-((4-(3-(3-isopropyl-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(4-methoxyphenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)-pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(4-methoxyphenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-methoxyethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(2,3,5,6-tetradeutero-4-(trideuteromethyl)phenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2,5,8,11-tetraoxatridecan-13-yl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)-pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-methoxyethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)-5,6,7,8-tetrahydronaphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(2,4-dimethoxyphenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(3-((dimethylphosphoryl)methyl)phenyl)-1H-pyrazol-5-yl)ureido)    naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(3-((dimethylphosphoryl)methyl)phenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;-   3-((4-((4-(3-(3-(tert-butyl)-1-(4-methoxy-2-methylphenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide,    and pharmaceutically acceptable salts thereof.

Thus in one embodiment the compound of the invention is3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamideor a pharmaceutically acceptable salt thereof.

In an alternative embodiment, there is provided a compound of formula(I), (Ib1), (Ib2), (Ic1), (Ic2), (Id1), (Id2), (Ig1) or (Ig2) as definedabove, or a pharmaceutically acceptable salt thereof, wherein thecompound is not a compound of the formula:

or a pharmaceutically acceptable salt thereof, including all tautomersthereof.

In certain embodiments, there is provided a compound of formula (I),(Ib1), (Ib2), (Ic1), (Ic2), (Id1), (Id2), (Ig1) or (Ig2) as definedabove, or a pharmaceutically acceptable salt thereof, wherein thecompound is not:

-   3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;    and/or-   3-((4-((4-(3-(3-(tert-butyl)-1-(2,3,5,6-tetradeutero-4-(trideuteromethyl)phenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide,    or a pharmaceutically acceptable salt thereof.

The pharmaceutically acceptable acid addition salts of compounds offormula (I) are meant to comprise the therapeutically active non-toxicacid addition salts that the compounds of formula (I) are able to form.These pharmaceutically acceptable acid addition salts can convenientlybe obtained by treating the free base form with such appropriate acidsin a suitable solvent or mixture of solvents. Appropriate acidscomprise, for example, inorganic acids such as hydrohalic acids, e.g.hydrochloric or hydrobromic acid, sulfuric, nitric, phosphoric acids andthe like; or organic acids such as, for example, acetic, propanoic,hydroxyacetic, lactic, pyruvic, malonic, succinic, maleic, fumaric,malic, tartaric, citric, methanesulfonic, ethanesulfonic,benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic,p-aminosalicylic, pamoic acid and the like.

Conversely said salt forms can be converted by treatment with anappropriate base into the free base form.

Stereoisomers as employed herein refers to isomeric molecules that havethe same molecular formula and sequence of bonded atoms (constitution),but that differ only in the three-dimensional orientations of theiratoms in space. This contrasts with structural isomers, which share thesame molecular formula, but the bond connections and/or their orderdiffer(s) between different atoms/groups. In stereoisomers, the orderand bond connections of the constituent atoms remain the same, but theirorientation in space differs.

As employed herein below the definition of compounds of formula (I) isintended to include all tautomers of said compounds, and solvates ofsaid compounds (including solvates of salts of said compounds) unlessthe context specifically indicates otherwise. Examples of solvatesinclude hydrates.

The invention provided herein extends to prodrugs of the compound offormula (I), that is to say compounds which break down and/or aremetabolised in vivo to provide an active compound of formula (I).General examples of prodrugs include simple esters, and other esterssuch as mixed carbonate esters, carbamates, glycosides, ethers, acetalsand ketals.

In a further aspect of the invention there is provided one or moremetabolites of the compound of formula (I), in particular a metabolitethat retains one or more of the therapeutic activities of the compoundof formula (I). A metabolite, as employed herein, is a compound that isproduced in vivo from the metabolism of the compound of formula (I),such as, without limitation, oxidative metabolites and/or metabolitesgenerated, for example, from 0-dealkylation.

The compounds of the disclosure include those where the atom specifiedis a naturally occurring or non-naturally occurring isotope. In oneembodiment the isotope is a stable isotope. Thus the compounds of thedisclosure include, for example deuterium containing compounds and thelike.

The disclosure also extends to all polymorphic forms of the compoundsherein defined.

Generic routes by which compound examples of the invention may beconveniently prepared are summarized below. Those routes arespecifically exemplified for compounds of formula (I) in which R^(a) andR^(b), together with the C-atoms to which they are attached, form afused phenyl ring. However, compounds of formula (I) having otherdefinitions of R^(a) and R^(b) may be prepared by analogous routes.

Thus, for example, compounds of formula (I) may be obtained by a generalprocess (Scheme 1, Route A) whereby a naphthylamine precursorrepresented by Intermediate B is coupled with an activated,electrophilic derivative Intermediate A* prepared from the correspondingamine precursor, Intermediate A (G=H). The amine radical NR^(a)R^(b) incompounds of Intermediate B either comprise the group Y, as defined forcompounds of formula (I) above or a protected derivative of the same.The fragment LG₁ in Intermediate A* is a suitable leaving group such asan imidazolyl (C₃H₃N₂) or an aryloxy radical such as a phenoxy (C₆H₅O)group. It will be understood by persons skilled in the art that, in someinstances, the compound represented by Intermediate A* may be isolatedor in other cases may be a transient intermediate, that is not isolated,but generated in situ and used directly.

In the case wherein LG₁ is imidazolyl, compounds represented byIntermediate A* are obtained by reaction of the corresponding amine withan activating agent such as CDI in a non-polar aprotic solvent, such asDCM and are conveniently generated in situ at RT and then reactedwithout isolation with compounds represented by Intermediate B.

In the case wherein LG₁ is aryloxy the required activated amine may begenerated by treatment of the amine precursor with a suitablechloroformate, such as, for example, phenyl chloroformate, in thepresence of a base. In some instances it is advantageous to conduct theactivation process under Schotten-Baumann type conditions, that is usingan aqueous base, such as aq sodium carbonate under biphasic conditions.The activated amine derivatives represented by Intermediate A* whereinLG₁ is aryloxy, for example phenoxy, may thereby be generated optionallyin situ and then reacted without isolation with compounds represented byIntermediate B to provide compound examples of formula (I).

Compounds of formula (I) may include those in which the substituent Yincorporates one or more functional groups that have been protectedduring the coupling process and therefore require(s) subsequentdeprotection. An example of such a procedure is the removal of atert-butoxycarbonyl (Boc) group from a secondary amine, by treatmentwith an appropriate acid.

Alternatively, compound examples of formula (I) may be generated by anS_(N)Ar displacement reaction between an electrophilic heteroaryloxyfragment represented by Intermediate C, wherein LG₂ is a suitableleaving group, typically a halogen atom, for example chlorine, with ananiline component represented by Intermediate D (Scheme 2, Route B). Thereaction proceeds under acidic conditions, for example in the presenceof p-TSA and in a polar aprotic solvent such as THF and typically atelevated temperatures, for example at 70° C.

Optionally, compound examples of the invention may be prepared by ageneral synthetic process comprising of an amide bond forming reactionbetween a carboxylic acid derivative with an amine R^(a)R^(b)NH (Scheme3, Routes C₁ and C₂) whereby NR^(a)R^(b) comprises Y or a protectedderivative thereof, in which latter case the compounds of formula (I)are revealed following an appropriate deprotection step(s). The amidecoupling may be conducted on an alkyl ester represented by IntermediateE (R^(c)=alkyl), for example a methyl ester, with the amine, in thepresence of a trialkylaluminium, for example trimethylaluminium (Scheme3, Route C₁). The reaction is conveniently carried out in an aproticsolvent such as THF and at ambient or slightly elevated temperatures,typically RT to 40° C.

Alternatively the amide products of formula (I) may be derived from theparent carboxylic acids represented by Intermediate F (R^(c)═H) byreaction with the amine R^(a)R^(b)NH under the influence of an amide(peptide) coupling reagent, and in the presence a non-nucleophilic base(Scheme 3, Route C₂). An example of a reagent that is frequentlyemployed for these transformations is HATU and suitable bases includeDIPEA and N-methylmorpholine and the like. The amidation reaction istypically conducted in polar aprotic solvents such as THF and at ambienttemperature.

The above and other routes may be used to prepare the compound offormula (I). Thus, according to a further aspect of the invention, thereis provided a process for the preparation of a compound of formula (I)which process comprises:

(a) reaction of a compound of formula (II),

with a compound of formula (III)

wherein LG^(II) represents a suitable leaving group (e.g. imidazolyl,halo (such as chloro) or, particularly, aryloxy (such as phenoxy)) andone of Z¹ and Z² is a structural fragment of formula (IV)

wherein R and Q are as hereinbefore defined, and the other of Z¹ and Z²is a structural fragment of formula (V)

wherein R¹, R^(a), R^(b), X and Y are as hereinbefore defined (e.g. inone particular embodiment, Z¹ is a structural fragment of formula (IV)and Z² is a structural fragment of formula (V)), for example underconditions known to those skilled in the art, such as from ambienttemperature to about 80° C. (e.g. from 50 to 60° C.), optionally in thepresence of an amine base (e.g. a trialkylamine such asN,N-diisopropylethylamine or, particularly, triethylamine) and asuitable organic solvent (e.g. an aprotic solvent, such asdichloromethane or, particularly, an ester such as isopropyl acetate);(b) reaction of a compound of formula (VII

wherein R, R^(a), R^(b), Q and X are as hereinbefore defined and LG^(VI)represents a suitable leaving group (e.g. a halo group such as bromo or,particularly, chloro), with a compound of formula (VII),

wherein R¹ and Y are as hereinbefore defined, under conditions known tothose skilled in the art (e.g. as described in J. Am. Chem. Soc. 2011,133, 15686-15696), such as at elevated temperature (e.g. from 50 to 110°C., such as at about 60° C.) in the presence of a suitable organicsolvent (e.g. a polar aprotic solvent such as DMF, THF, 1,4-dioxane, ormixtures thereof) and, optionally, an acidic catalyst (e.g. a sulfonicacid such as para-toluenesulfonic acid);(c) reaction of a compound of formula (VIII),

with a compound of formula (III), wherein the compound of formula (III)and Z¹ and Z² are as hereinbefore defined, under conditions known tothose skilled in the art, for example at a temperature from ambient(e.g. 15 to 30° C.) to about 110° C. in the presence of a suitableorganic solvent (e.g. a polar aprotic solvent such as DMF, THF,1,4-dioxane, or mixtures thereof);(d) reaction of a compound of formula (IX),

wherein Z¹ is as defined above, with a suitable azide-forming agent(i.e. a suitable source of a leaving group and activated azide ion, suchas diphenyl phosphorazidate; see, for example, Tetrahedron 1974, 30,2151-2157) under conditions known to those skilled in the art, such asat sub-ambient to ambient temperature (e.g. from an initial temperatureof about −5 to 5° C. to ambient temperature post-reaction) in thepresence of an amine base (e.g. triethylamine or a sterically hinderedbase such as N,N-diisopropylethylamine) and a suitable organic solvent(e.g. a polar aprotic solvent such as DMF, THF, 1,4-dioxane, or mixturesthereof), which reaction is followed, without isolation, by thermalrearrangement (e.g. under heating) of the intermediate acyl azide (offormula Z¹—C(O)—N₃) e.g. at ambient temperature (such as from 15 to 30°C.) to provide, in situ, a compound of formula (VIII), which compound isthen reacted with a compound of formula (III), as defined above, toprovide the compound of formula (I);(e) reaction of a compound of formula (X)

wherein R, R¹, R^(a), R^(b), Q and X are as hereinbefore defined andR^(X) represents H or C₁₋₄ alkyl, with a compound of formula (XI)

wherein R² and R³ are as hereinbefore defined, under conditions known tothose skilled in the art, for example

-   -   when R^(X) represents H, reaction in the presence of a suitable        solvent, a base (e.g. triethylamine or        N,N-diisopropylethylamine) and an amide (peptide) coupling        reagent, such as HATU, CDI, N,N′-dicyclohexylcarbodiimide,        N,N′-diisopropylcarbodiimide BOP or PyBOP, optionally in        combination with an activated ester-forming agent such as HOBt        or 1-hydroxy-7-azabenzotriazole,    -   when R^(X) represents H, conversion of the carboxylic acid to an        acid halide (e.g. by reaction with a halogenating agent such as        thionyl chloride), followed by reaction with the compound of        formula (XI) in the presence of a suitable solvent and a base        (e.g. triethylamine or N,N-diisopropylethylamine), or    -   when R^(X) represents C₁₋₄ alkyl (e.g. methyl), reaction in the        presence of a trialkylaluminium (e.g. trimethylaluminium) and an        aprotic solvent (e.g. THF); or        (f) deprotection of a protected derivative of a compound of        formula (I), under conditions known to those skilled in the art,        wherein the protected derivative bears a protecting group on an        O- or N-atom of the compound of formula (I) (and, for the        avoidance of doubt, a protected derivative of one compound of        formula (I) may or may not represent another compound of formula        I).

Compounds represented by Intermediate A are either commerciallyavailable, or may be prepared by synthetic approaches that are wellestablished in the art. For example compounds of this general structuremay be prepared by condensation of the appropriate hydrazine, optionallyin the form of a protected derivative thereof or a suitable salt, withthe relevant ketonitrile (Scheme 4). An example of an appropriate saltis a hydrochloride salt, and a suitable protective group for thistransformation is an acid labile carbamate, for example a Boc group(R^(d)=tert-Bu) that is readily removed under the cyclisation conditions

to generate the parent hydrazine in situ. The condensation/cyclisationreaction is suitably conducted in a polar protic solvent such as ethanoland in the presence of a strong acid for example concentratedhydrochloric acid and at elevated temperatures, typically at reflux.

In some instances it may be advantageous to prepare such intermediatesby one or other alternative methodologies, as best suits theavailability of starting materials and/or the functionality representedin the compounds and/or the need to protect one or more of them, duringthe synthetic processes in question or in subsequent transformations.For example compounds represented by Intermediate A may also be accessedvia a copper (I) mediated coupling reaction between a 1H-pyrazol-5-amineand a suitable arene Q-LG₃ in which Q is an optionally substitutedaromatic nucleus as defined for compounds of formula (I) and LG₃ is ahalide such as an iodine atom (Scheme 5). The reaction is convenientlyconducted in an

aprotic non-polar solvent such as toleune, employing a copper (I) saltas the catalyst, for example copper (I) iodide and in the presence of acopper co-ordinating ligand such asN¹,N²-dimethylcyclohexane-1,2-diamine and in the presence of a base, forexample potassium carbonate and typically at elevated temperature forexample at reflux.

It will be evident to those skilled in the art that it may beadvantageous to convert one intermediate described herein into anotherexample of the same by one or more transformations that are well knownand precedented and thereby gain access to additional compounds of theinvention. As an example of such a process those compounds representedby Intermediate A wherein Q is a phenyl ring substituted with an alkoxygroup (OR^(e) wherein R^(e) is alkyl), such as a methoxy group, may beconverted into the corresponding phenol by an O-dealkylation reaction(Scheme 6). This type of transformation may be effected with a borontrihalide, for example boron tribromide, in a non-polar, aprotic solventsuch as DCM, at reduced temperatures for example at −5 to 0° C.

A further demonstration of the conversion of one intermediate, intoanother compound of the same generic type is provided by thefunctionalisation of the phenol examples of Intermediate A describedhereinabove. For example intermediates of this composition can beconveniently alkylated on the phenolic oxygen by reaction with an alkylhalide, for example with a simple alkyl bromide. Alternatively, thephenol products may be reacted with a functionalised alkyl halide, forexample with a nitrogen mustard, that is, with a salt of a2-haloethylamine of formula R^(f)(CH₂)₂LG₄, wherein LG₄ is a halogensuch as a chlorine and R^(f) is selected such that O(CH₂)₂R^(f) isallowable by the definition of Q in compounds of formula (I)

or is a suitably protected derivative thereof (Scheme 7). An example ofa salt of a 2-haloethylamine that could be used in O-alkylations of thiskind is 4-(2-chloroethyl)morpholine hydrochloride. Reaction of this kindare usefully undertaken in polar non protic solvents such asacetonitrile or DMF and in the presence of a base such as potassiumcarbonate and with heating if necessary.

In some instances it may be advantageous to effect the O-alkylationunder Mitsunobu conditions, by interaction of the phenol with thecorresponding alcohol R^(f)(CH₂)₂OH in the presence of a triarylphosphine such as triphenyphosphine, together with a suitablediazodicarboxylate coupling reagent, for example diisopropyldiazene-1,2-dicarboxylate. Such reactions are typically carried out innon-polar, aprotic solvents such as THF at reduced to ambienttemperatures, for example at −50° C. to RT.

Other examples of Intermediate A may be prepared by interconversion ofsubstituents on the phenyl, pyridyl or thienyl ring of group Q. Forexample, examples of Intermediate A bearing the substituent—(CH₂)₁₋₂P(O)R′R″ may be obtained by coupling of equivalent compoundsbearing the substituent —(CH₂)₁₋₂Hal, where Hal is a leaving group suchas chloro, bromo or iodo, with a compound of the formula H—P(O)R′R″. Thereaction may be performed, for example, by heating in a polar aproticsolvent (e.g. DMF) in the presence of a palladium-containing catalyst(e.g. a Pd(II) catalytic species, such as Pd(II) acetate, optionally inthe presence of a bidentate phosphine ligand such as4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos))Alternatively, the coupling reaction may be with a compound of formula(C₁₋₄ alkyl)-O—PR′R″, using Arbuzov-type conditions (WO 2010/141406;Bioorg. Med. Chem. Lett. 2009, 19, 2053-2058), with compounds having the—(CH₂)₁₋₂Hal substituent mentioned above. (Compounds of formula (C₁₋₄alkyl)-O—PR′R″ are typically made in situ by reaction of thecorresponding chlorophosphine (Cl—PR′R″) with a C₁₋₄ alkyl alcohol inthe presence or a base (e.g. diisopropylethylamine), or with an alkalimetal salt of a C₁₋₄ alkyl alcohol.)

Compounds represented by Intermediate B may be obtained from S_(N)Ardisplacement reactions between electrophilic aryloxy naphthylaminesrepresented by Intermediate G, wherein LG₂ is a suitable leaving groupsuch as a halogen atom, for example chlorine, with an aniline componentrepresented by Intermediate D (Scheme 8). The coupling reaction may beundertaken on the free naphthylamine (G₁=H) or optionally, in order tocontrol chemoselectivity and thereby enhance efficiency, upon aprotected derivative thereof

Intermediate G(P)

(G₁=protective group). The reaction proceeds under acidic conditions,for example in the presence of p-TSA and in a polar aprotic solvent suchas THF and typically at elevated temperatures, for example at 70° C. Inthose instances in which a protective group has been employed theproducts represented by Intermediate B are subsequently revealed by asuitable deprotection step(s). For example a carbamate, such as a Bocgroup, may be used to protect the naphthylamine nitrogen (G₁=tert-BuO₂C)during the S_(N)Ar coupling reaction and afterwards removed by treatmentwith a strong acid, for example with TFA.

The synthetic processes cited hereinabove (Routes C₁ and C₂, Scheme 3)may likewise be exploited to access compounds represented byIntermediate B (Scheme 9). Thus examples of Intermediate B may beprepared by reaction of an activated derivative of a carboxylic acidrepresented by Intermediate J (R^(c)=G₁=H) or a protected derivativethereof Intermediate J(P) (G₁=protective group) with an amineR^(a)R^(b)NH, whereby NR^(a)R^(b) comprises Y or a protected derivativethereof. Alternatively the interconversion may be undertaken on an esterIntermediate H (R^(c)=alkyl, G₁=H) or a protected derivative thereofIntermediate H(P) (R^(c)=alkyl, G₁=protective group) with an amineR^(a)R^(b)NH in the presence of a trialkyl aluminium, as alreadydescribed. A suitable protective group for these transformations is aurethane

derivative (G₁=R^(h)O₂C) in which case the desired anilines (G₁=H)represented by Intermediate B are obtained following an appropriatedeprotection procedure. An example of a urethane protective group whichis suitable for this purpose is a Boc group (G₁=tert-BuO₂C), which canbe removed following the amidation reaction by treatment with acid.

The ester and acid precursors represented by Intermediates E and F areobtainable by use of the same or analogous procedures, to thosedisclosed hereinabove (Scheme 1), that provide compound examples of thepresent invention. In this manner Intermediates E and F are convenientlyobtained by the reaction of Intermediates H and J respectively with theactivated aminopyrazole derivatives Intermediates A* (Scheme 10). Itwill be evident to those skilled in the art that the esters:Intermediates H and E may be readily transformed into the correspondingcarboxylic acids: Intermediates J and F by hydrolysis under suitableacidic or basic conditions. For example this conversion can be effectedby saponification, using a base such as lithium hydroxide, in a proticsolvent or mixture of solvents, for example THF and water and atmodestly elevated temperatures, typically RT to 40° C.

The precursors represented by Intermediate G are conveniently preparedby an S_(N)Ar displacement reaction between 4-aminonaphthalen-1-ol,either in the form of a salt or a suitable, protected derivative and anelectrophilic heteroaromatic (Scheme 11), for example a dihaloheteroaromatic wherein the leaving groups LG₂ and LG₅ are both halogenatoms,

such as chlorine. A suitable protective group for this transformation isa Boc group (G₁=tert-BuO₂C) which may be retained, in order to controlchemoselectivity, during one or more subsequent transformations, such asthose described hereinabove (Schemes 8 and 9). The displacement step isconveniently carried out in a polar, aprotic solvent such asacetonitrile and in the presence of a hindered base, typified by DBU andat reduced temperature, for example at 0° C.

Those compounds represented by Intermediates H and J were assembled byanalogous synthetic procedures to those already described above (Scheme8) for the preparation of Intermediates B by substituting anilino acidsor anilino esters represented by Intermediate K in place of IntermediateD (Scheme 12). In a similar manner the acid mediated S_(N)Ar couplingmay be conducted on the free naphthylamine Intermediate G (G₁=H) oroptionally, using a protected derivative of the same, Intermediate G(P)(G₁=protective group), to maintain the desired chemoselectivity in thisand/or subsequent transformations. The S_(N)Ar coupling is suitablycarried out in a polar non protic solvent, for example THF or IPA or DMFand in the presence of an acid catalyst such as p-TSA or TFA and mostusually at elevated temperatures, typically at 60-70° C. In alternativeprocedures, a Buchwald-Hartwig amination may be performed, for exampleat elevated temperature (e.g. 20 to 120° C.) using a palladium catalyst(e.g. a combination of Pd₂dba₃ and BINAP) and base (e.g. Cs₂CO₃). Ininstances where the phenyl group of the aniline bears a substituent thatis sensitive to palladium-catalysed reactions (e.g. an ethynyl group),then Intermediate K may be replaced by Intermediate K*, in which thesensitive (e.g. ethynyl) group is in protected form (e.g. for ethynyl,in trialkylsilyl-protected form, such as in triisopropylsilane-protectedform). The protective group may then be removed under conditions knownto those skilled in the art. For example, a triisopropylsilyl group canbe removed with a source of fluoride ion, e.g., tetrabutylammoniumfluoride (TBAF) or caesium fluoride.

The known aniline components represented by Intermediate K were eitherprocured from commercial sources or prepared according to publishedprocedures. Novel examples of Intermediate D and Intermediate Kdisclosed herein were synthesised from commercially available startingmaterials using functional group interconversions that are wellestablished in the art (Scheme 13). For example, the (leaving) group LG₆may be displaced with a desired R¹ group via an S_(N)Ar reaction ortransition metal-catalysed coupling. In some instances the desiredanilines are readily obtainable from appropriately substituted, aminobenzoic acids (R^(c)=G₂=H) and/or amino benzoic acid esters (R^(c)=alkylG₂=H) that may be optionally N-protected (G₂=PG) to ensure thatsubsequent reactions can be conducted effectively. Transposition of thesubstituent R^(h) into a group R¹ as defined for compounds of formula(I), provides compounds represented by Intermediate K which may behydrolysed and subjected to an amide coupling reaction to furnishexamples of Intermediate D, after removal, where employed, of thenitrogen protective group.

Additional examples of Intermediate D are readily made from commerciallyavailable (protected) amino or nitro benzoic acids that are substitutedwith a suitable (leaving) group LG₆, such as a halogen, for examplefluorine or, particularly, bromine. Compounds of this composition may beconverted into the examples of the desired anilines by a series ofreactions comprising of an amide coupling, followed by an S_(N)Ardisplacement reaction and reduction of the nitro group into an amine.Alternatively, for preparing examples of Intermediate D in which R¹represents —C≡C—(C₁₋₄ alkylene)₀₋₁-H, the (protected) amino or nitrobenzoic acids may undergo a Sonogashira coupling reaction and (forprotected amino or nitro compounds) either deprotection of the aminogroup or reduction of the nitro group into an amine. Alternatively, theSonogashira coupling reaction may be performed before the amide coupling(Scheme 14). In either alternative, when the alkynyl moiety introducedby way of the Sonogashira coupling reaction is ethynyl, it is introducedin protected form (e.g. in trialkylsilyl-protected form, through use ofan ethynyltrialkylsilane, such as ethynyltriisopropylsilane). In suchinstances, the protective (e.g. trialkylsilyl) group may be removedunder conditions known to those skilled in the art. For example, atriisopropylsilyl group can be removed with a source of fluoride ion,e.g., tetrabutylammonium fluoride (TBAF) or caesium fluoride. For theSonogashira coupling reaction, group LG₆ is suitably a halo group suchas chloro, iodo or, particularly, bromo.

Compounds of formula (I) may alternatively be obtained by coupling ofIntermediate B to an pyrazole-5-isocyanate compound, Intermediate L. Inthis route, Intermediate L may, for example, be conveniently preparedvia a copper (II)-mediated Chan-Lam reaction (see, for example:Tetrahedron Lett. 1998, 39, 2941-2944), wherein an ester of a suitablepyrazole-5-carboxylic acid is coupled to an aryl- or heteroaryl-boronicacids. The resulting N-aryl pyrazole acid ester is saponified to yieldthe corresponding carboxylic acid (Intermediate M), which acid isconverted to an acyl azide (e.g. using source of a leaving group andactivated azide ion, such as diphenyl phosphorazidate (DPPA); see, forexample, Tetrahedron 1974, 30, 2151-2157)) before undergoing a Curtisrearrangement to yield Intermediate L.

As exemplified in Scheme 15 above, examples of Intermediate M(P) may beprepared by metal-catalysed coupling of pyrazole acid esters to suitablecompounds containing aromatic group Q. However, other examples ofIntermediate M(P) may be prepared by further elaboration of compounds soobtained. For example, when Q is phenyl, a chloro, bromo or iodosubstituent on the phenyl group in a compound of Intermediate M(P) maybe displaced by cross-coupling with a variety of nucleophilic groups,such as a dialkylphosphine oxide (Scheme 16). The cross-couplingtypically employs a palladium-containing catalyst (e.g. a Pd(II)catalytic species, such as Pd(II) acetate, optionally in the presence ofa bidentate phosphine ligand such as4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (Xantphos); see, forexample, WO 2009/143389).

It will be evident to those skilled in the art that in some cases it istechnically advantageous to use alternative protective groups and/or toconduct the transformations described above in a similar manner but in adifferent order, so as to improve the overall efficiency of thesynthetic processes.

Protective groups and the means for their removal are described in“Protective Groups in Organic Synthesis”, by Theodora W. Greene andPeter G. M. Wuts, published by John Wiley & Sons Inc; 4th Rev Ed., 2006,ISBN-10: 0471697540.

Novel intermediates as described herein form an aspect of the invention.In this respect, further aspects of the invention relate to:

(i) a compound of formula (III) as hereinbefore defined, wherein Z²represents a structural fragment of formula (V), as hereinbeforedefined, or a salt or protected derivative thereof; and(ii) Intermediate D, as hereinbefore defined (i.e. a compound of formula(VII), as hereinbefore defined) or a salt or protected derivativethereof.

Protected derivatives of Intermediates B and D include amides or,particularly, carbamates of those compounds. For example, thoseprotected derivatives include compounds in which a H-atom of the NH₂group is replaced by:

-   -   R′—C(O)—, wherein R′ is H, C₁₋₈ alkyl, phenyl or benzyl, which        latter two groups are optionally substituted by one or more        groups selected from halo, hydroxy, methyl and methoxy; or    -   R″—O—C(O)—, wherein R″ is tert-butyl, phenyl, benzyl or        fluorenyl, which latter three groups are optionally substituted        by one or more groups selected from halo, hydroxy, methyl and        methoxy.

The compounds of formula (I) are p38 MAP kinase inhibitors (especiallyof the alpha subtype) and in one aspect the compounds are useful in thetreatment of inflammatory diseases, for example COPD and/or asthma.

Surprisingly, in at least some embodiments, the compounds of formula (I)exhibit a long duration of action and/or persistence of action.

In one embodiment the compounds of formula (I) do not strongly inhibit,or bind to GSK 3α, for example they have an IC₅₀ value against GSK 3α of1500 nM or greater; such as 2,000, 3,000, 4,000, 5,000, 6,000, 7,000,8,000, 9,000 or 10,000 nM or greater.

Persistence of action as used herein is related to the dissociation rateor dissociation constant of the compound from the target (such as areceptor). A low dissociation rate may lead to persistence.

A low dissociation rate in combination with a high association ratetends to provide potent therapeutic entities.

The compounds of formula (I) are expected to be potent in vivo.

Typically, the prior art compounds developed to date have been intendedfor oral administration. This strategy involves optimizing thepharmacokinetic profile of drug substances in order to achieve anadequate duration of action. In this manner a sufficiently high drugconcentration is established and maintained between doses to providesustained clinical benefit. The inevitable consequence of this approachis that all bodily tissues, and especially the liver and the gut, arelikely to be exposed to supra-therapeutically active concentrations ofthe drug, whether or not they are adversely affected by the diseasebeing treated.

An alternative strategy is to design treatment paradigms in which thedrug is dosed directly to the inflamed organ, that is, to exploittopical administration. Whilst this approach is not suitable fortreating all chronic inflammatory diseases, it has been exploited inlung disorders, such as asthma and COPD; in skin diseases, for exampleagainst atopic dermatitis and psoriasis; for nasal conditions, typifiedby allergic rhinitis; and in gastrointestinal diseases, such asulcerative colitis and Crohn's disease and inflammatory diseases of theeye, such as uveitis.

In topical therapy, one way in which efficacy can be achieved is by theuse of a drug that has a sustained duration of action and is retained inthe relevant organ, thereby minimizing the risk of systemic toxicity.Alternatively, in some cases, a formulation can be developed thatgenerates a “reservoir” of the active drug which is available to sustainits desired effects. The first approach is exemplified by theanticholinergic drug tiotropium (Spiriva). This compound is administeredtopically to the lung as a treatment for COPD, and has an exceptionallyhigh affinity for its target receptor resulting in a very slow off rateand consequently displays a sustained duration of action.

In one aspect of the disclosure the compounds of formula (I) isparticularly suitable for topical delivery, such as topical delivery tothe lungs, in particular for the treatment of respiratory disease, forexample chronic respiratory diseases such as COPD and/or asthma.

In one embodiment the compounds of formula (I) is suitable forsensitizing patients to treatment with a corticosteroid who have becomerefractory to such treatment regimens.

The compounds of formula (I) may have antiviral properties (such asthose described in WO 2011/070368 and/or WO 2011/070369), for examplethe ability to prevent the infection of cells (such as respiratoryepithelial cells) with a picornavirus, in particular a rhinovirus,influenza or respiratory syncytial virus.

Thus, in view of their kinase inhibition profiles, the compounds arethought to be antiviral agents, in particular suitable for theprevention, treatment or amelioration of picornavirus infections, suchas rhinovirus infection, influenza or respiratory syncytial virus.

In one embodiment the compounds of formula (I) are able to reduceinflammation induced by viral infection, such as rhinovirus infectionand in particular viral infections that result in the release ofcytokines such as IL-8, especially in vivo. This activity may, forexample, be tested in vitro employing a rhinovirus induced IL-8 assay.

In one embodiment the compounds of formula (I) are able to reduce ICAM1expression induced by rhinovirus, especially in vivo. ICAM1 is thereceptor mechanism used by so-called major groove rhinovirus serotypesto infect cells. This activity may be measured, for example by a methoddescribed herein.

It is expected that the above properties render the compounds of formula(I) particularly suitable for use in the treatment (includingprophylaxis) of exacerbations of inflammatory diseases, in particularviral exacerbations, or in the treatment of viral infections, inpatients with one or more chronic conditions such as congestive heartfailure, COPD, asthma, diabetes, cancer and/or in immunosuppressedpatients, for example post-organ transplant. Such use may be incombination with anti-viral agents such as zanamivir, oseltamivir (forexample oseltamivir phosphate) peramivir or laninamivir.

In general, the compounds of formula (I) may be useful in the treatmentof one or more conditions having an inflammatory component which,suitably, may be treated by topical or local therapy.

In particular, the compounds of formula (I) may be useful in thetreatment of one or more respiratory disorders including COPD (includingchronic bronchitis and emphysema), asthma, pediatric asthma, cysticfibrosis, sarcoidosis, idiopathic pulmonary fibrosis, allergic rhinitis,rhinitis and sinusitis, especially asthma, or COPD (including chronicbronchitis and emphysema).

The compounds of formula (I) may be useful in the treatment of eyediseases or disorders including keratoconjunctivitis sicca (dry eye),allergic conjunctivitis, conjunctivitis, diabetic retinopathy, macularoedema (including wet macular oedema and dry macular oedema),post-operative cataract inflammation or, particularly, uveitis(including posterior, anterior and pan uveitis) (e.g. eye diseases ordisorders including allergic conjunctivitis, conjunctivitis, diabeticretinopathy, macular oedema (including wet macular oedema and drymacular oedema), post-operative cataract inflammation or, particularly,uveitis (including posterior, anterior and pan uveitis)).

The compounds of formula (I) may be useful in the treatment of skindiseases or disorders including allergic dermatitis, contact dermatitis,atopic dermatitis or psoriasis.

The compounds of formula (I) may be useful in the treatment ofgastrointestinal diseases or disorders including ulcerative colitis orCrohn's disease.

The compounds of formula (I) may be useful in the treatment of jointdiseases or disorders including rheumatoid arthritis or osteoarthritisand particularly inflamed joints secondary to such conditions.

The compounds of formula (I) may be useful in the treatment of cancersincluding cancer of the stomach and in the inhibition of the growth andmetastasis of tumours including non-small cell lung carcinoma, gastriccarcinoma, colorectal carcinomas and malignant melanoma.

It is also expected that the compounds of formula (I) may be useful inthe treatment of certain other conditions including periodontitis,gingivitis and pharyngitis.

Compounds of formula (I) may also re-sensitise the patient's conditionto treatment with a corticosteroid, when the patient's condition hasbecome refractory to the same.

Furthermore, the present invention provides a pharmaceutical compositioncomprising a compound according to the disclosure optionally incombination with one or more pharmaceutically acceptable diluents orcarriers.

Diluents and carriers may include those suitable for parenteral, oral,topical, mucosal and rectal administration.

The present invention also provides a process for preparing such apharmaceutical composition (for example a pharmaceutical composition forparenteral, oral, topical, mucosal or rectal administration) whichcomprising mixing the ingredients.

As mentioned above, such compositions may be prepared e.g. forparenteral, subcutaneous, intramuscular, intravenous, intra-articular orperi-articular administration, particularly in the form of liquidsolutions or suspensions; for oral administration, particularly in theform of tablets or capsules; for topical e.g. pulmonary or intranasaladministration, particularly in the form of powders, nasal drops oraerosols and transdermal administration; for mucosal administration e.g.to buccal, sublingual or vaginal mucosa, and for rectal administratione.g. in the form of a suppository.

The compositions may conveniently be administered in unit dosage formand may be prepared by any of the methods well-known in thepharmaceutical art, for example as described in Remington'sPharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa.,(1985). Formulations for parenteral administration may contain asexcipients sterile water or saline, alkylene glycols such as propyleneglycol, polyalkylene glycols such as polyethylene glycol, oils ofvegetable origin, hydrogenated naphthalenes and the like. Formulationsfor nasal administration may be solid and may contain excipients, forexample, lactose or dextran, or may be aqueous or oily solutions for usein the form of nasal drops or metered sprays. For buccal administrationtypical excipients include sugars, calcium stearate, magnesium stearate,pregelatinated starch, and the like.

Compositions suitable for oral administration may comprise one or morephysiologically compatible carriers and/or excipients and may be insolid or liquid form. Tablets and capsules may be prepared with bindingagents, for example, syrup, acacia, gelatin, sorbitol, tragacanth, orpoly-vinylpyrollidone; fillers, such as lactose, sucrose, corn starch,calcium phosphate, sorbitol, or glycine; lubricants, such as magnesiumstearate, talc, polyethylene glycol, or silica; and surfactants, such assodium lauryl sulfate. Liquid compositions may contain conventionaladditives such as suspending agents, for example sorbitol syrup, methylcellulose, sugar syrup, gelatin, carboxymethyl-cellulose, or ediblefats; emulsifying agents such as lecithin, or acacia; vegetable oilssuch as almond oil, coconut oil, cod liver oil, or peanut oil;preservatives such as butylated hydroxyanisole (BHA) and butylatedhydroxytoluene (BHT). Liquid compositions may be encapsulated in, forexample, gelatin to provide a unit dosage form.

Solid oral dosage forms include tablets, two-piece hard shell capsulesand soft elastic gelatin (SEG) capsules.

A dry shell formulation typically comprises of about 40% to 60% w/wconcentration of gelatin, about a 20% to 30% concentration ofplasticizer (such as glycerin, sorbitol or propylene glycol) and about a30% to 40% concentration of water. Other materials such aspreservatives, dyes, opacifiers and flavours also may be present. Theliquid fill material comprises a solid drug that has been dissolved,solubilized or dispersed (with suspending agents such as beeswax,hydrogenated castor oil or polyethylene glycol 4000) or a liquid drug invehicles or combinations of vehicles such as mineral oil, vegetableoils, triglycerides, glycols, polyols and surface-active agents.

Suitably a compound of formula (I) is administered topically to thelung, eye or bowel. Hence we provide according to the invention apharmaceutical composition comprising a compound of the disclosureoptionally in combination with one or more topically acceptable diluentsor carriers.

Topical administration to the lung may be achieved by use of an aerosolformulation. Aerosol formulations typically comprise the activeingredient suspended or dissolved in a suitable aerosol propellant, suchas a chlorofluorocarbon (CFC) or a hydrofluorocarbon (HFC).

Suitable CFC propellants include trichloromonofluoromethane (propellant11), dichlorotetrafluoromethane (propellant 114), anddichlorodifluoromethane (propellant 12). Suitable HFC propellantsinclude tetrafluoroethane (HFC-134a) and heptafluoropropane (HFC-227).The propellant typically comprises 40% to 99.5% e.g. 40% to 90% byweight of the total inhalation composition. The formulation may compriseexcipients including co-solvents (e.g. ethanol) and surfactants (e.g.lecithin, sorbitan trioleate and the like). Aerosol formulations arepackaged in canisters and a suitable dose is delivered by means of ametering valve (e.g. as supplied by Bespak, Valois or 3M).

Topical administration to the lung may also be achieved by use of anon-pressurised formulation such as an aqueous solution or suspension.This may be administered by means of a nebuliser. Topical administrationto the lung may also be achieved by use of a dry-powder formulation. Adry powder formulation will contain the compound of the disclosure infinely divided form, typically with a mass mean aerodynamic diameter(MMAD) of 1-10 μm.

The formulation will typically contain a topically acceptable diluentsuch as lactose, usually of large particle size e.g. an MMAD of 100 μmor more. Examples of dry powder delivery systems include SPINHALER,DISKHALER, TURBOHALER, DISKUS and CLICKHALER.

The compounds of the present invention (i.e. compounds of formula (I),(Ia1), (Ia2), (Ib1), (Ib2), (Ic1), (Ic2), (Id1), (Id2), (Ie1), (Ie2),(If1), (If2), (Ig1), (Ig2), (Ih1) or (Ih2), as defined above, orpharmaceutically acceptable salts thereof) may also be administeredrectally, for example in the form of suppositories or enemas, whichinclude aqueous or oily solutions as well as suspensions and emulsions.Such compositions are prepared following standard procedures, well knownby those skilled in the art. For example, suppositories can be preparedby mixing the active ingredient with a conventional suppository basesuch as cocoa butter or other glycerides. In this case, the drug ismixed with a suitable non-irritating excipient which is solid atordinary temperatures but liquid at the rectal temperature and willtherefore melt in the rectum to release the drug. Such materials arecocoa butter and polyethylene glycols.

Generally, for compositions intended to be administered topically to theeye in the form of eye drops or eye ointments, the total amount of theinhibitor will be about 0.0001 to less than 4.0% (w/w).

Preferably, for topical ocular administration, the compositionsadministered according to the present invention will be formulated assolutions, suspensions, emulsions and other dosage forms. Aqueoussolutions are generally preferred, based on ease of formulation, as wellas a patient's ability to administer such compositions easily by meansof instilling one to two drops of the solutions in the affected eyes.However, the compositions may also be suspensions, viscous orsemi-viscous gels, or other types of solid or semi-solid compositions.Suspensions may be preferred for compounds that are sparingly soluble inwater.

An alternative for administration to the eye is intravitreal injectionof a solution or suspension of the compound of the present invention. Inaddition, the compound of the present invention may also be introducedby means of ocular implants or inserts.

The compositions administered according to the present invention mayalso include various other ingredients, including, but not limited to,tonicity agents, buffers, surfactants, stabilizing polymer,preservatives, co-solvents and viscosity building agents. Preferredpharmaceutical compositions of the present invention include theinhibitor with a tonicity agent and a buffer. The pharmaceuticalcompositions of the present invention may further optionally include asurfactant and/or a palliative agent and/or a stabilizing polymer.

Various tonicity agents may be employed to adjust the tonicity of thecomposition, preferably to that of natural tears for ophthalmiccompositions. For example, sodium chloride, potassium chloride,magnesium chloride, calcium chloride, simple sugars such as dextrose,fructose, galactose, and/or simply polyols such as the sugar alcoholsmannitol, sorbitol, xylitol, lactitol, isomaltitol, maltitol, andhydrogenated starch hydrolysates may be added to the composition toapproximate physiological tonicity. Such an amount of tonicity agentwill vary, depending on the particular agent to be added. In general,however, the compositions will have a tonicity agent in an amountsufficient to cause the final composition to have an ophthalmicallyacceptable osmolality (generally about 150-450 mOsm, preferably 250-350mOsm and most preferably at approximately 290 mOsm). In general, thetonicity agents of the invention will be present in the range of 2 to 4%w/w. Preferred tonicity agents of the invention include the simplesugars or the sugar alcohols, such as D-mannitol.

An appropriate buffer system (e.g., sodium phosphate, sodium acetate,sodium citrate, sodium borate or boric acid) may be added to thecompositions to prevent pH drift under storage conditions. Theparticular concentration will vary, depending on the agent employed.Preferably however, the buffer will be chosen to maintain a target pHwithin the range of pH 5 to 8, and more preferably to a target pH of pH5 to 7.

Surfactants may optionally be employed to deliver higher concentrationsof inhibitor. The surfactants function to solubilize the inhibitor andstabilise colloid dispersion, such as micellar solution, microemulsion,emulsion and suspension. Examples of surfactants which may optionally beused include polysorbate, poloxamer, polyosyl 40 stearate, polyoxylcastor oil, tyloxapol, triton, and sorbitan monolaurate. Preferredsurfactants to be employed in the invention have ahydrophile/lipophile/balance “HLB” in the range of 12.4 to 13.2 and areacceptable for ophthalmic use, such as TritonX114 and tyloxapol.

Additional agents that may be added to the ophthalmic compositions ofthe present invention are demulcents which function as a stabilisingpolymer. The stabilizing polymer should be an ionic/charged example withprecedence for topical ocular use, more specifically, a polymer thatcarries negative charge on its surface that can exhibit a zeta-potentialof (−)10-50 mV for physical stability and capable of making a dispersionin water (i.e. water soluble). A preferred stabilising polymer of theinvention would be polyelectrolyte, or polyelectrolytes if more thanone, from the family of cross-linked polyacrylates, such as carbomersand Pemulen®, specifically Carbomer 974p (polyacrylic acid), at 0.1-0.5%w/w.

Other compounds may also be added to the ophthalmic compositions of thepresent invention to increase the viscosity of the carrier. Examples ofviscosity enhancing agents include, but are not limited to:polysaccharides, such as hyaluronic acid and its salts, chondroitinsulfate and its salts, dextrans, various polymers of the cellulosefamily; vinyl polymers; and acrylic acid polymers.

Topical ophthalmic products are typically packaged in multidose form.Preservatives are thus required to prevent microbial contaminationduring use. Suitable preservatives include: benzalkonium chloride,chlorobutanol, benzododecinium bromide, methyl paraben, propyl paraben,phenylethyl alcohol, edentate disodium, sorbic acid, polyquaternium-1,or other agents known to those skilled in the art. Such preservativesare typically employed at a level of from 0.001 to 1.0% w/v. Unit dosecompositions of the present invention will be sterile, but typicallyunpreserved. Such compositions, therefore, generally will not containpreservatives.

The medical practitioner, or other skilled person, will be able todetermine a suitable dosage for the compounds of the invention, andhence the amount of the compound of the invention that should beincluded in any particular pharmaceutical formulation (whether in unitdosage form or otherwise).

A compound of formula (I) has therapeutic activity. In a further aspect,the present invention provides a compound of the disclosure for use as amedicament. Thus, in a further aspect, the present invention provides acompound as described herein for use in the treatment of one or more ofthe above mentioned conditions.

In one embodiment a dry powder formulation according the presentdisclosure comprises magnesium or calcium stearate. Such formulationsmay have superior chemical and/or physical stability especially whensuch formulations also contain lactose.

In a further aspect, the present invention provides use of a compound asdescribed herein for the manufacture of a medicament for the treatmentof one or more of the above mentioned conditions.

In a further aspect, the present invention provides a method oftreatment of one or more of the above mentioned conditions whichcomprises administering to a subject an effective amount of a compoundof the disclosure or a pharmaceutical composition comprising thecompound.

The word “treatment” is intended to embrace prophylaxis as well astherapeutic treatment. Treatment of conditions or disorders alsoembraces treatment of exacerbations thereof.

A compound of the disclosure may also be administered in combinationwith one or more other active ingredients e.g. active ingredientssuitable for treating the above mentioned conditions.

For example, possible combinations for treatment of respiratorydisorders include combinations with steroids (e.g. budesonide,beclomethasone dipropionate, fluticasone propionate, mometasone furoate,fluticasone furoate), beta agonists (e.g. terbutaline, salbutamol,salmeterol, formoterol), xanthines (e.g. theophylline), anticholinergics(e.g. ipratropium or tiotropium, for example as the bromide) andanti-viral agents (e.g. zanamivir, oseltamivir, for example as thephosphate, peramivir and laninamivir).

Further, for the treatment of gastrointestinal disorders (such asCrohn's disease or ulcerative colitis), possible combinations includecombinations with, for example, one or more agents selected from thelist comprising:

-   -   5-aminosalicylic acid, or a prodrug thereof (such as        sulfasalazine, olsalazine or bisalazide);    -   corticosteroids (e.g. prednisolone, methylprednisolone, or        budesonide);    -   immunosuppressants (e.g. cyclosporin, tacrolimus, methotrexate,        azathioprine or 6-mercaptopurine);    -   anti-TNFα antibodies (e.g., infliximab, adalimumab, certolizumab        pegol or golimumab);    -   anti-IL12/IL23 antibodies (e.g., ustekinumab) or small molecule        IL12/IL23 inhibitors (e.g., apilimod);    -   Anti-α4β7 antibodies (e.g., vedolizumab);    -   MAdCAM-1 blockers (e.g., PF-00547659);    -   antibodies against the cell adhesion molecule α4-integrin (e.g.,        natalizumab);    -   antibodies against the IL2 receptor a subunit (e.g., daclizumab        or basiliximab);    -   JAK3 inhibitors (e.g., tofacitinib or R348);    -   Syk inhibitors and prodrugs thereof (e.g., fostamatinib and        R-406);    -   Phosphodiesterase-4 inhibitors (e.g., tetomilast);    -   HMPL-004;    -   probiotics;    -   Dersalazine;    -   semapimod/CPSI-2364; and    -   protein kinase C inhibitors (e.g. AEB-071).

For the treatment of eye disorders (such as keratoconjunctivitis siccaor uveitis), possible combinations include combinations with, forexample, one or more agents selected from the list comprising:

-   -   corticosteroids (e.g. dexamethasone, prednisolone, triamcinolone        acetonide, difluprednate or fluocinolone acetonide);    -   immunosuppressants (e.g. cyclosporin, voclosporin, azathioprine,        methotrexate, mycophenolate mofetil or tacrolimus);    -   anti-TNFα antibodies (e.g., infliximab, adalimumab, certolizumab        pegol, ESBA-105 or golimumab);    -   anti-IL-17A antibodies (e.g., secukinumab);    -   mTOR inhibitors (e.g., sirolimus);    -   VGX-1027;    -   JAK3 inhibitors (e.g., tofacitinib or R348); and    -   protein kinase C inhibitors (e.g. AEB-071).

Hence another aspect of the invention provides a compound of formula (I)in combination with one or more further active ingredients, for exampleone or more active ingredients described above.

Similarly, another aspect of the invention provides a combinationproduct comprising:

-   (A) a compound of the present invention (i.e. a compound of formula    (I), (Ia1), (Ia2), (Ib1), (Ib2), (Ic1), (Ic2), (Id1), (Id2), (Ie1),    (Ie2), (If1), (If2), (Ig1), (Ig2), (Ih1) or (Ih2), as defined above,    or a pharmaceutically acceptable salt thereof); and-   (B) another therapeutic agent,    wherein each of components (A) and (B) is formulated in admixture    with a pharmaceutically-acceptable adjuvant, diluent or carrier.

In this aspect of the invention, the combination product may be either asingle (combination) pharmaceutical formulation or a kit-of-parts.

Thus, this aspect of the invention encompasses a pharmaceuticalformulation including a compound of the present invention and anothertherapeutic agent, in admixture with a pharmaceutically acceptableadjuvant, diluent or carrier (which formulation is hereinafter referredto as a “combined preparation”).

It also encompasses a kit of parts comprising components:

-   (i) a pharmaceutical formulation including a compound of the present    invention in admixture with a pharmaceutically acceptable adjuvant,    diluent or carrier; and-   (ii) a pharmaceutical formulation including another therapeutic    agent, in admixture with a pharmaceutically-acceptable adjuvant,    diluent or carrier,    which components (i) and (ii) are each provided in a form that is    suitable for administration in conjunction with the other.

Component (i) of the kit of parts is thus component (A) above inadmixture with a pharmaceutically acceptable adjuvant, diluent orcarrier. Similarly, component (ii) is component (B) above in admixturewith a pharmaceutically acceptable adjuvant, diluent or carrier.

The other therapeutic agent (i.e. component (B) above) may be, forexample, any of the agents mentioned above in connection with thetreatment of respiratory, gastrointestinal and eye disorders.

The combination product (either a combined preparation or kit-of-parts)of this aspect of the invention may be used in the treatment orprevention of an inflammatory disease (e.g. the inflammatory diseasesmentioned above, such as:

-   -   respiratory disorders including COPD (including chronic        bronchitis and emphysema), asthma, pediatric asthma, cystic        fibrosis, sarcoidosis, idiopathic pulmonary fibrosis, allergic        rhinitis, rhinitis and sinusitis, especially asthma, or COPD        (including chronic bronchitis and emphysema);    -   eye diseases or disorders including allergic conjunctivitis,        conjunctivitis, keratoconjunctivitis sicca (dry eye), glaucoma,        diabetic retinopathy, macular oedema (including diabetic macular        oedema), central retinal vein occlusion (CRVO), dry and/or wet        age related macular degeneration (AMD), post-operative cataract        inflammation or, particularly, uveitis (including posterior,        anterior and pan uveitis), corneal graft and limbal cell        transplant rejection;    -   skin diseases or disorders including allergic dermatitis,        contact dermatitis, atopic dermatitis or psoriasis; and    -   gastrointestinal diseases or disorders including gluten        sensitive enteropathy (coeliac disease), eosinophilic        esophagitis, intestinal graft versus host disease or,        particularly, ulcerative colitis or Crohn's disease.

The aspects of the invention described herein (e.g. the above-mentionedcompound, combinations, methods and uses) may have the advantage that,in the treatment of the conditions described herein, they may be moreconvenient for the physician and/or patient than, be more efficaciousthan, be less toxic than, be longer acting than, have better selectivityover, have a broader range of activity than, be more potent than,produce fewer side effects than, have a better pharmacokinetic and/orpharmacodynamic profile than, have more suitable solid state morphologythan, have better stability than, or may have other usefulpharmacological properties over, similar compounds, combinations,methods (treatments) or uses known in the prior art for use in thetreatment of those conditions or otherwise.

Relative to compounds of the prior art, the compounds of formula (I) mayadditionally (or alternatively):

-   -   exhibit properties that are particularly suited to topical/local        administration (e.g. following topical/local administration, the        generation of high target tissue concentrations but low plasma        concentrations of the compounds of formula (I) and/or rapid        clearance of the compounds of formula (I) from plasma);    -   have a reduced risk of extravascular exposure following        intravenous administration (e.g. due to a low volume of        distribution for the compounds of formula (I));    -   exhibit superior potency with respect to selected kinases (e.g.        Syk and/or a panel of kinases, such as Syk, Src and p38 MAPKα);    -   exhibit reduced β-catenin induction and/or inhibition of mitosis        in cells;    -   exhibit no or less time-dependent inhibition of members of the        cytochrome P450 superfamily; and/or    -   produce less problematic (e.g. less toxic) metabolites, e.g.        following administration to a patient.

EXPERIMENTAL SECTION

Abbreviations used herein are defined below (Table 1). Any abbreviationsnot defined are intended to convey their generally accepted meaning.

TABLE 1 Abbreviations AcOH glacial acetic acid aq aqueous ATPadenosine-5′-triphosphate BALF bronchoalveolar lavage fluid br broad BSAbovine serum albumin CatCart ® catalytic cartridge CDI1,1-carbonyl-diimidazole COPD chronic obstructive pulmonary diseasec-Src cellular sarc(oma) kinase d doublet DCM dichloromethane DMEMDulbecco's Modified Eagle Medium DMSO dimethyl sulfoxide DSS dextransodium sulphate d-U937 cells PMA differentiated U-937 cells (ES⁺)electrospray ionization, positive mode Et ethyl EtOAc ethyl acetate FCSfoetal calf serum FRET fluorescence resonance energy transfer GRglucocorticoid receptor GSK3α glycogen synthase kinase 3α HBEC primaryhuman bronchial epithelial cells hr hour(s) HRP horseradish peroxidiseHRV human rhinovirus IBD inflammatory bowel disease ICAM-1inter-cellular adhesion molecule 1 IL-8 interleukin 8 JNK c-JunN-terminal kinase LPS lipopolysaccharide (M + H)⁺ protonated molecularion MAPK mitogen-activated protein kinase MAPKAP-K2 mitogen-activatedprotein kinase-activated protein kinase-2 Me methyl MeCN acetonitrileMeOH methanol MHz megahertz MMAD mass median aerodynamic diameter MOImultiplicity of infection min minute(s) MPO myeloperoxidase MTT3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide m/z:mass-to-charge ratio NMR nuclear magnetic resonance (spectroscopy) NTNot tested PBMC peripheral blood mononuclear cell PBS phosphate bufferedsaline PG protective group Ph phenyl PHA phytohaemagglutinin PMA phorbolmyristate acetate p-TSA 4-methylbenzenesulfonic acid q quartet RT roomtemperature RP HPLC reverse phase high performance liquid chromatographyRSV respiratory syncytial virus s singlet sat saturated SCX solidsupported cation exchange (resin) SDS sodium dodecyl sulphate S_(N)Arnucleophilic aromatic substitution Syk spleen tyrosine kinase t tripletT3P 1-propanephosphonic acid cyclic anhydride TBDMStert-butyldimethylsilyl TCID₅₀ 50% tissue culture infectious dose THFtetrahydrofuran TMB 3,3′,5,5′-tetramethylbenzidine TNBS2,4,6-trinitrobenzenesuifonic acid TNFα tumor necrosis factor alpha WBwashing buffer

General Procedures

All starting materials and solvents were obtained either from commercialsources or prepared according to the literature citation. Unlessotherwise stated all reactions were stirred. Organic solutions wereroutinely dried over anhydrous magnesium sulfate. Hydrogenations wereperformed on a Thales H-cube flow reactor under the conditions stated.

Column chromatography was performed on pre-packed silica (230-400 mesh,40-63 μm) cartridges using the amount indicated. SCX was purchased fromSupelco and treated with 1M hydrochloric acid prior to use. Unlessstated otherwise the reaction mixture to be purified was first dilutedwith MeOH and made acidic with a few drops of AcOH. This solution wasloaded directly onto the SCX and washed with MeOH. The desired materialwas then eluted by washing with 0.7 M NH₃ in MeOH.

Preparative Reverse Phase High Performance Liquid Chromatography

Agilent Scalar column C18, 5 μm (21.2×50 mm), flow rate 28 mL min⁻¹eluting with a H₂O-MeCN gradient containing 0.1% v/v formic acid over 10min using UV detection at 215 and 254 nm. Gradient information: 0.0-0.5min; 95% H₂O-5% MeCN; 0.5-7.0 min; ramped from 95% H₂O-5% MeCN to 5%H₂O-95% MeCN; 7.0-7.9 min; held at 5% H₂O-95% MeCN; 7.9-8.0 min;returned to 95% H₂O-5% MeCN; 8.0-10.0 min; held at 95% H₂O-5% MeCN.

Analytical Methods

Reverse Phase High Performance Liquid Chromatography

Method 1:

Agilent Scalar column C18, 5 μm (4.6×50 mm) or Waters XBridge C18, 5 μm(4.6×50 mm) flow rate 2.5 mL min⁻¹ eluted with a H₂O-MeCN gradientcontaining either 0.1% v/v formic acid (Method 1 acidic) or NH₃ (Method1 basic) over 7 min employing UV detection at 215 and 254 nm. Gradientinformation: 0.0-0.1 min, 95% H₂O-5% MeCN; 0.1-5.0 min, ramped from 95%H₂O-5% MeCN to 5% H₂O-95% MeCN; 5.0-5.5 min, held at 5% H₂O-95% MeCN;5.5-5.6 min, held at 5% H₂O-95% MeCN, flow rate increased to 3.5 mLmin⁻¹; 5.6-6.6 min, held at 5% H₂O-95% MeCN, flow rate 3.5 mL min⁻¹;6.6-6.75 min, returned to 95% H₂O-5% MeCN, flow rate 3.5 mL min⁻¹;6.75-6.9 min, held at 95% H₂O-5% MeCN, flow rate 3.5 mL·min⁻¹; 6.9-7.0min, held at 95% H₂O-5% MeCN, flow rate reduced to 2.5 mL min⁻¹.

Method 2:

Agilent Extend C18 column, 1.8 μm (4.6×30 mm) at 40° C.; flow rate2.5-4.5 mL min⁻¹ eluted with a H₂O-MeCN gradient containing either 0.1%v/v formic acid (Method 2 acidic) or NH₃ (Method 2 basic) over 4 minemploying UV detection at 254 nm. Gradient information: 0-3.00 min,ramped from 95% H₂O-5% MeCN to 5% H₂O-95% MeCN; 3.00-3.01 min, held at5% H₂O-95% MeCN, flow rate increased to 4.5 mL min⁻¹; 3.01 3.50 min,held at 5% H₂O-95% MeCN; 3.50-3.60 min, returned to 95% H₂O-5% MeCN,flow rate reduced to 3.50 mL min⁻¹; 3.60-3.90 min, held at 95% H₂O-5%MeCN; 3.90-4.00 min, held at 95% H₂O-5% MeCN, flow rate reduced to 2.5mL min⁻¹.

Method 3:

Waters Xselect CSH C18 3.5 μm (4.6×50 mm) flow rate 2.5 mL min⁻¹ elutedwith a H₂O-MeCN gradient containing 0.1% v/v formic acid over 7 minemploying UV detection at 215 and 254 nm. Gradient information: 0.0-0.1min, 95% H₂O-5% MeCN; 0.1-5.0 min, ramped from 95% H₂O-5% MeCN to 5%H₂O-95% MeCN; 5.0-5.5 min, held at 5% H₂O-95% MeCN; 5.5-5.6 min, held at5% H₂O-95% MeCN, flow rate increased to 3.5 mL min⁻¹; 5.6-6.6 min, heldat 5% H₂O-95% MeCN, flow rate 3.5 mL min⁻¹; 6.6-6.75 min, returned to95% H₂O-5% MeCN, flow rate 3.5 mL min⁻¹; 6.75-6.9 min, held at 95%H₂O-5% MeCN, flow rate 3.5 mL·min⁻¹; 6.9-7.0 min, held at 95% H₂O-5%MeCN, flow rate reduced to 2.5 mL min⁻¹.

Method 4:

Waters Xselect CSH C18 3.5 μm (4.6×50 mm); flow rate 2.5-4.5 mL min⁻¹eluted with a H₂O-MeCN gradient containing 0.1% v/v formic acid over 4min employing UV detection at 254 nm. Gradient information: 0-3.00 min,ramped from 95% H₂O-5% MeCN to 5% H₂O-95% MeCN; 3.00-3.01 min, held at5% H₂O-95% MeCN, flow rate increased to 4.5 mL min⁻¹; 3.01 3.50 min,held at 5% H₂O-95% MeCN; 3.50-3.60 min, returned to 95% H₂O-5% MeCN,flow rate reduced to 3.50 mL min⁻¹; 3.60-3.90 min, held at 95% H₂O-5%MeCN; 3.90-4.00 min, held at 95% H₂O-5% MeCN, flow rate reduced to 2.5mL min⁻¹.

¹H NMR Spectroscopy

¹H NMR spectra were acquired on a Bruker Avance III spectrometer at 400MHz using residual undeuterated solvent as reference and unlessspecified otherwise were run in DMSO-d₆.

Those intermediates, used to prepare compound examples of the invention,that have been previously disclosed were obtained using the procedurescontained in the references cited below (Table 2). Additionalintermediates were prepared by the representative synthetic processesdescribed herein.

TABLE 2 Compound Intermediates Name, LCMS Data No. Structure andReference A1

3-tert-butyl-1- p-tolyl-pyrazol- 5-amine. R^(t) 2.46 min (Method 1basic); m/z 230 (M + H)⁺, (ES⁺). Cirillo, P. F. et al., WO 2000/43384,27 Jul. 2000. A1*

phenyl (3- (tert-butyl)- 1-(p-tolyl)-1H- pyrazol-5-yl) carbamate LCMSm/z 350 (M + H)⁺ (ES⁺); 348 (M − H)⁻ (ES⁻) Kapadia, S. R. et al., U.S.Pat. No. 6,492,529, 10 Dec. 2002. A2

3-isopropyl-1-(p- tolyl)-1H-pyrazol-5- amine. R^(t) 3.14 min (Method 1,acidic, X-Select); m/z 216 (M + H)⁺, (ES⁺). Ito, K. et al., WO2010/067130, 17 Jun. 2010 A3

3-tert-butyl-1-(4- methoxyphenyl)- 1H-pyrazol- 5-amine. R^(t) 1.32 min(Method 2, acidic); m/z 246 (M + H)⁺, (ES⁺). Mathias, J. P. et al., US2006/0035922, 10 Aug. 2005. A3*

phenyl (3-(tert- butyl)-1-(4- methoxyphenyl)- 1H-pyrazol-5-yl) carbamateLCMS m/z 366 (M + H)⁺ (ES⁺); 364 (M − H)⁻ (ES⁻) Abraham, S. et al., WO2009/117080, 24 Sep. 2009. A4

3-tert-butyl-1-(2,3, 5,6-tetradeutero- 4-(trideuteromethyl) phenyl)-1H-pyrazole-5-amine m/z 237 (M + H)⁺, (ES⁺). Ito, K. et al., WO2010/067130, 17 Jun. 2010 G1

4-((2-chloropyridin- 4-yl)oxy)naphthalen- 1-amine. R^(t) 3.13 min(Method 3); m/z 271/273 (M + H)⁺, (ES⁺). Ito, K. et al., WO 2010/112936,07 Oct. 2010 G2

4-((2-chloro- pyrimidin- 4-yl)oxy) naphthalen-1- amine. R^(t) 1.80 min(Method 2, acidic); m/z 272/ 274 (M + H)⁺, (ES⁺). Cirillo, P. F. et al.,WO 2002/92576, 21 Nov. 2000. G2(P)

tert-butyl (4-((2- chloropyrimidin- 4-yl)oxy) naphthalen-1-yl)carbamate. R^(t) 2.43 min (Method 2, acidic); m/z 372/ 374 (M + H)⁺,(ES⁺). Ito, K. et al., WO 2010/067130, 17 Jun. 2010

Intermediate A4*Phenyl(3-(tert-butyl)-1-(2,3,5,6-tetradeutero-4-(trideuteromethyl)-phenyl)-1H-pyrazol-5-yl)carbamate

Phenyl chloroformate (580 μL, 4.62 mmol) was added to a stirred mixtureof Intermediate A4 (1 g, 4.23 mmol) and NaHCO₃ (0.711 g, 8.46 mmol) inDCM (20 mL) and THF (10 mL). The mixture was stirred for 3 h thenpartitioned between DCM (200 mL) and water (100 mL). The organic layerwas separated, washed with brine, dried (MgSO₄), filtered and evaporatedunder reduced pressure. The residue was triturated with ether/isohexane,filtered and dried to afford the sub-title compound (1.335 g) LCMS m/z357 (M+H)⁺ (ES⁺); 355 (M−H)⁻ (ES⁻)

Intermediate A53-(tert-Butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-amine

DPPA (0.550 mL, 2.55 mmol) was added to a stirred solution ofIntermediate M1 (500 mg, 1.740 mmol) and Et₃N (0.4 mL, 2.87 mmol) intert-butanol (10 ml) under N₂ then heated to reflux for 18 h. Themixture was cooled, water (75 mL) added and extracted with ethyl acetate(3×50 mL). The combined organic phases were washed with saturated brine(50 mL), dried (MgSO₄) and concentrated under reduced pressure. Thecrude product was purified by chromatography on the Companion (40 gcolumn, 25-100% DCM:iso-hexane) to affordtert-butyl(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)carbamate(459 mg) as a pale orange oil.

LCMS m/z 359 (M+H)⁺ (ES⁺); 357 (M−H)⁻ (ES⁻)

TFA (1 mL, 12.98 mmol) was added to a stirred solution of(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)carbamateobtained immediately above (459 mg, 1.280 mmol) in DCM (5 mL) at rt for3 h. The mixture was concentrated under reduced pressure and the residuewas redissolved in ethyl acetate (25 mL). The organic solution waswashed with saturated NaHCO₃ solution (2×25 mL), saturated brine (25mL), dried (MgSO₄) and concentrated under reduced pressure to yieldIntermediate A5 (335 mg).

LCMS m/z 259 (M+H)⁺ (ES⁺)

Intermediate A5*Phenyl(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)carbamate

Phenyl chloroformate (175 μL, 1.395 mmol) was added to a suspension ofIntermediate A5 (335 mg, 1.297 mmol) and NaHCO₃ (220 mg, 2.62 mmol) inTHF (4 mL) and DCM (4 mL). The mixture was stirred at rt for 2 h. Themixture was diluted with DCM (20 mL) and washed with water (25 mL). Theorganic phase was washed with saturated brine (20 mL), dried (MgSO₄) andconcentrated under reduced pressure. The residue was recrystallised incyclohexane to afford the sub-title compound (345 mg) as a white solid.¹H NMR (CDCl₃) 400 MHz, δ: 7.44-7.31 (m, 4H), 7.29-7.22 (m, 1H),7.21-7.11 (m, 2H), 7.03-6.90 (m, 1H), 6.90-6.76 (m, 2H), 6.56-6.36 (m,1H), 3.04 (s, 6H), 1.37 (s, 9H).

LCMS m/z 379 (M+H)⁺ (ES⁺)

Intermediate B13-((4-((4-Aminonaphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide

Pd₂dba₃ (22 mg, 0.024 mmol) and BINAP (30 mg, 0.048 mmol) were stirredin 1,4-dioxane (1 mL) for 10 minutes under N₂. In a separate vessel,purged with N₂, caesium carbonate (455 mg, 1.396 mmol), Intermediate D3(291 mg, 0.930 mmol) and Intermediate G1(P) (345 mg, 0.930 mmol) werestirred in 1,4-dioxane (5 mL). The catalyst solution was added to themain reaction mixture and the whole was heated to 90° C. for 18 h. Uponcooling, the mixture was diluted with water (40 mL) and extracted withethyl acetate (3×25 mL). The combined organic phases were washed withsaturated brine (15 mL), dried (MgSO₄) and concentrated under reducedpressure. The crude product was purified by chromatography on theCompanion (40 g column, 0-50% acetone/ethyl acetate) to affordtert-butyl(4-((2-((3-methoxy-5-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)carbamoyl)phenyl)amino)pyridin-4-yl)oxy)-naphthalen-1-yl)carbamate(Intermediate B1(P), 320 mg) as a sticky orange oil.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.37 (s, 1H), 9.09 (s, 1H), 8.35 (t, 1H),8.17-8.05 (m, 2H), 7.83 (d, 1H), 7.67-7.46 (m, 5H), 7.35 (d, 1H), 6.88(s, 1H), 6.57 (dd, 1H), 6.09 (d, 1H), 3.74 (s, 3H), 3.58-3.44 (m, 8H),3.44-3.34 (m, 4H), 3.20 (s, 3H), 1.52 (s, 9H).

LCMS m/z 647 (M+H)⁺ (ES⁺); 645 (M−H)⁻ (ES⁻)

A solution of Intermediate B1(P) (320 mg, 0.495 mmol) in DCM (1 mL) wastreated with TFA (1000 μL, 12.98 mmol) and stirred at rt for 3 h. Themixture was diluted with water (10 mL) and DCM (10 mL). The mixture wasneutralised with saturated NaHCO₃ and passed through a phase separationcartridge. The organic phase was dried (MgSO₄) and concentrated to giveIntermediate B1 (270 mg) as a brown gum.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.00 (s, 1H), 8.34 (dd, 1H), 8.20-8.10 (m,1H), 8.05 (d, 1H), 7.67-7.60 (m, 1H), 7.59-7.55 (m, 1H), 7.52-7.47 (m,1H), 7.47-7.41 (m, 2H), 7.10 (d, 1H), 6.89-6.84 (m, 1H), 6.71 (d, 1H),6.51 (dd, 1H), 6.05 (d, 1H), 5.83 (s, 2H), 3.73 (S, 3H), 3.58-3.45 (m,8H), 3.45-3.35 (m, 4H), 3.21 (s, 3H).

LCMS m/z 547 (M+H)⁺ (ES⁺)

Intermediate B23-((4-((4-Aminonaphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide

Method 1

T3P, 50% w/w in EtOAc (54.0 ml, 91 mmol) was added to a solutionIntermediate J1(P) (30 g, 60.4 mmol),2-(2-(2-methoxyethoxy)ethoxy)ethanamine (11.83 g, 72.5 mmol), and TEA(20 mL, 143 mmol) in DMF (400 mL). The mixture was stirred at rt for 18h. The mixture was diluted with water (700 mL) and saturated sodiumhydrogen carbonate solution (500 mL) and the mixture was extracted withethyl acetate (3×400 mL). The combined organic phases were washed with20% brine (3×500 mL), saturated brine (3×500 mL), dried (MgSO₄),filtered and concentrated in vacuo. The crude product was purified intwo batches by chromatography on the companion (330 g column, 1-5% MeOHin DCM) to affordtert-butyl(4-((2-((3-ethynyl-5-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)carbamoyl)phenyl)amino)-pyrimidin-4-yl)oxy)naphthalen-1-yl)carbamate(Intermediate B2(P), 24.4 g) as a pale yellow foam.

¹H NMR (400 MHz, DMSO-d6) δ: 9.74 (s, 1H), 9.31 (s, 1H), 8.44-8.47 (m,2H), 8.11 (s, 1H), 8.10 (d, 1H), 7.91 (s, 1H), 7.82 (d, 1H), 7.54-7.63(m, 3H), 7.46 (s, 1H), 7.42 (d, 1H), 6.58 (d, 1H), 4.15 (s, 1H),3.49-3.53 (m, 8H), 3.36-3.41 (m, 4H), 3.21 (s, 3H), 1.52 (s, 9H).

LCMS m/z 642 (M+H)⁺ (ES⁺); 640 (M−H)⁻ (ES⁻)

Trifluoroacetic acid (30 ml, 389 mmol) was added dropwise to a stirredsolution of Intermediate B2(P) (12.0 g, 18.70 mmol) in DCM (200 mL). Thereaction was stirred at rt for 3 h. The reaction was concentrated invacuo and the residue partitioned between DCM (300 mL) and saturatedNaHCO₃ solution (400 mL). The aqueous phase was separated and extractedwith DCM (200 mL). The combined organics were dried (MgSO₄), filteredand concentrated in vacuo to give a beige foam. The crude product waspurified by chromatography on the Companion (220 g column, 1-5% MeOH inDCM) to afford Intermediate B2 (9.0 g) as a pale pink foam.

¹H NMR (400 MHz, DMSO-d6) δ: 9.74 (s, 1H), 8.46 (t, 1H), 8.36 (d, 1H),8.12-8.14 (m, 1H), 8.07 (s, 1H), 7.94 (s, 1H), 7.62-7.64 (m, 1H),7.41-7.46 (m, 3H), 7.15 (d, 1H), 6.70 (d, 1H), 6.38 (d, 1H), 5.76 (s,2H), 4.18 (s, 1H), 3.49-3.53 (m, 8H), 3.36-3.41 (m, 4H), 3.21 (s, 3H).LCMS m/z 542 (M+H)⁺ (ES⁺); 540 (M−H)⁻ (ES⁻)

Method 2

A mixture of Intermediate D2 (155.76 g of approx. 90% purity, 458 mmol),Intermediate G2(P) (166.6 g, 448 mmol) and para-toluenesulfonic acid(14.7 g, 85 mmol) in THF (2.5 L) was heated at reflux for 6 h. Thereaction mixture was then allowed to cool overnight to provide a darkbrown mixture having a “jelly” consistency. Solvent was evaporated toprovide a viscous, tacky material. This was dissolved in mixture ofethyl acetate and sodium bicarbonate (aqueous). The aqueous and organiclayers were separated and the aqueous layer was re-extracted with ethylacetate. The combined ethyl acetate layers were washed with brine (×2)before being dried (MgSO₄), filtered through a silica plug, reduced (bysolvent evaporation) to approx 10% volume and then poured onto a largeexcess of n-hexane. The resulting mixture was stirred for 6-8 h toprovide a solid that was isolated by filtration, washed with n-hexaneand then dried to afford a pale brown solid (Intermediate B2(P), 223 g,77.5%).

Washing of the silica plug with acetone yielded a further 42 g (14.6%)of mixture of Intermediate B2(P) and Intermediate B2, as determined byTLC.

The two solid materials comprising or containing Intermediate B2(P) werecombined and then used in the next stage without any furtherpurification.

A mixture of Intermediate B2(P) (265 g, 413 mmol), trifluoroacetic acid(575.62 g, 5.048 mol) and dichloromethane (2 L) was stirred together for3 h. Analysis by TLC (ethyl acetate) showed complete consumption ofstarting material. The solvent was reduced by evaporation and theresulting residue dissolved in ethyl acetate. The organic solution wascautiously washed with sodium bicarbonate (saturated, aq.) before beingdried (MgSO₄), filtered and then concentrated in vacuo to affordIntermediate B2 as a black gum/foam (200 g, 89.4%) that had ¹H NMR andLCMS data essentially identical to those of the material obtained viaMethod 1 above, and which gum/foam was used in the next step withoutfurther purification.

Intermediate B33-((4-((4-Aminonaphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2,5,8,11-tetraoxatridecan-13-yl)benzamide

T3P (50% w/w in EtOAc, 1.80 ml, 3.02 mmol) was added to a solution ofIntermediate J1(P) (1 g, 2.014 mmol), 2,5,8,11-tetraoxatridecan-13-amine(0.501 g, 2.417 mmol), and Et₃N (0.70 mL, 5.02 mmol) in DMF (15 mL). Thereaction was stirred at rt for 18 h. The mixture was diluted with water(200 mL) and saturated aqueous NaHCO₃ solution (100 mL) and the aqueousphase extracted with EtOAc (3×100 mL). The combined organic phases werewashed with 20% brine (3×100 mL), saturated brine (3×100 mL), then dried(MgSO₄), filtered and concentrated in vacuo affording a brown foam. Thecrude product was purified by chromatography on silica gel (40 g column,1-5% MeOH in DCM) to afford the sub-title compoundtert-butyl(4-((2-((3-((2,5,8,11-tetraoxatridecan-13-yl)carbamoyl)-5-ethynylphenyl)-amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)carbamate(Intermediate B3(P), 940 mg) as an off-white foam.

LCMS m/z 686 (M+H)⁺ (ES⁺); 684 (M−H)⁻ (ES⁻)

Trifluoroacetic acid (2000 μL, 26.0 mmol) was added dropwise to astirred solution of Intermediate B3(P) (940 mg, 1.371 mmol) in DCM (20ml). The reaction was stirred at rt overnight. The reaction wasconcentrated in vacuo and the residue partitioned between DCM (30 mL)and saturated aqueous NaHCO₃ solution (100 mL). The aqueous phase wasextracted with DCM (20 mL). The combined organics were dried (MgSO₄),filtered and concentrated in vacuo onto silica gel. The crude productwas purified by chromatography (40 g column, 3-5% MeOH in DCM) to affordthe sub-title compound (670 mg) as a pale yellow oil which becamefoam-like on drying at 40° C. under vacuum for 2 h.

¹H NMR (400 MHz, DMSO-d6) δ: 9.74 (s, 1H), 8.46 (t, 1H), 8.36 (d, 1H),8.12-8.14 (m, 1H), 8.07 (s, 1H), 7.94 (s, 1H), 7.62-7.64 (m, 1H),7.41-7.46 (m, 3H), 7.14 (d, 1H), 6.70 (d, 1H), 6.37 (d, 1H), 5.76 (s,2H), 4.17 (s, 1H), 3.47-3.53 (m, 12H), 3.36-3.41 (m, 4H), 3.22 (s, 3H).

LCMS m/z 586 (M+H)⁺ (ES⁺); 584 (M−H)⁻ (ES⁻)

Intermediate B43-((4-((4-Aminonaphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-methoxyethoxy)ethyl)benzamide

HATU (842 mg, 2.215 mmol) was added to a stirred solution ofIntermediate J1(P) (1000 mg, 2.014 mmol), 2-(2-methoxyethoxy)ethanamine(360 mg, 3.02 mmol) and Hunig's Base (1000 μL, 5.73 mmol) in DMF (10 mL)at rt. The mixture was stirred for 3 h then added to a vigorouslystirred solution of 0.1 M hydrogen chloride (200 mL). The resultingprecipitate was collected by filtration and washed with water (20 mL) toyield the crude product as a tan solid. The solid was purified bychromatography on silica gel (40 g column, 0-20% Acetone/EtOAc) toaffordtert-butyl(4-((2-((3-ethynyl-5-((2-(2-methoxyethoxy)ethyl)-carbamoyl)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)carbamate(Intermediate B4(P), 895 mg) as a yellow foam.

¹H NMR (400 MHz, DMSO-d6) δ 9.75 (s, 1H), 9.31 (s, 1H), 8.53-8.40 (m,1H), 8.45 (d, 1H), 8.17-8.06 (m, 2H), 7.95-7.87 (m, 1H), 7.87-7.77 (m,1H), 7.65-7.52 (m, 3H), 7.46 (d, 1H), 7.42 (d, 1H), 6.57 (d, 1H), 4.15(s, 1H), 3.58-3.47 (m, 4H), 3.47-3.35 (m, 4H), 3.23 (s, 3H), 1.52 (s,9H).

LCMS m/z 598 (M+H)⁺ (ES⁺); 596 (M−H)⁻ (ES⁻)

Trifluoroacetic acid (1.000 mL, 12.98 mmol) was added dropwise to astirred solution of Intermediate B4(P) (0.895 g, 1.498 mmol) in DCM (8mL). The reaction was stirred at rt for 18 h. The solvents wereevaporated and the residue partitioned between EtOAc (50 mL) and sat.NaHCO₃ soln. (50 ml), the organic phase was washed with saturated brine(50 ml). The organics were bulked, dried, filtered and evaporated togive a pale brown foam. The foam was purified by chromatography onsilica gel (12 g column, EtOAc) to afford Intermediate B4 (700 mg) as alight beige foam.

¹H NMR (400 MHz, DMSO-d6) δ 9.74 (s, 1H), 8.45 (dd, 1H), 8.36 (d, 1H),8.17-8.10 (m, 1H), 8.10-8.03 (m, 1H), 7.97-7.90 (m, 1H), 7.67-7.59 (m,1H), 7.48-7.38 (m, 3H), 7.15 (d, 1H), 6.70 (d, 1H), 6.37 (d, 1H),5.81-5.71 (m, 2H), 4.18 (s, 1H), 3.57-3.47 (m, 4H), 3.47-3.34 (m, 4H),3.23 (s, 3H).

LCMS m/z 498 (M+H)⁺ (ES⁺); 496 (M−H)⁻ (ES⁻)

Intermediate B53-((4-((4-Aminonaphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide

HATU (500 mg, 1.315 mmol) was added to a stirred solution ofIntermediate J2(P) (500 mg, 1.009 mmol),2-(2-(2-methoxyethoxy)ethoxy)ethanamine (277 mg, 1.695 mmol) and Et₃N(250 μL, 1.796 mmol) in DMF (10 mL). The mixture was stirred at rt for18 h. The mixture was diluted with ethyl acetate (50 mL) and washed withwater (50 mL), 20% brine (3×50 mL) and saturated brine (50 mL). Theorganic phase was dried (MgSO₄), filtered and concentrated under reducedpressure. The crude product was purified by chromatography on theCompanion (40 g column, EtOAc) to affordtert-butyl(4-((2-((3-ethynyl-5-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)carbamoyl)phenyl)amino)pyridin-4-yl)oxy)naphthalen-1-yl)carbamate(Intermediate B5(P), 580 mg) as a tan foam.

LCMS m/z 641 (M+H)⁺ (ES⁺); 639 (M−H)⁻ (ES⁻)

TFA (1 mL, 12.98 mmol) was added to a solution of Intermediate B5(P)(580 mg, 0.905 mmol) in DCM (5 mL) at rt and stirred overnight. Thevolatiles were removed under reduced pressure and the residue wasredissolved in DCM (20 mL). The organic phase was washed with saturatedNaHCO₃ solution (20 mL), dried (MgSO₄) and concentrated under reducedpressure to yield Intermediate B5 (475 mg).

LCMS m/z 541 (M+H)⁺ (ES⁺); 539 (M−H)⁻ (ES⁻)

Intermediate B63-((4-((4-Aminonaphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide

HATU (425 mg, 1.118 mmol) was added to a stirred solution ofIntermediate J3(P) (500 mg, 0.995 mmol),2-(2-(2-methoxyethoxy)ethoxy)ethanamine (250 mg, 1.532 mmol) and Hunig'sBase (500 μl, 2.86 mmol) in DMF (10 ml) at rt. The mixture was stirredfor 1 h then partitioned between 10% aq brine (100 ml) and EtOAc (100ml). The organic layer was washed with sat aq NaHCO₃ soln (50 ml), 0.5MHCl (50 ml), 10% aq brine (50 ml), dried (MgSO₄), filtered andevaporated under reduced pressure to affordtert-butyl(4-((2-((3-methoxy-5-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)carbamoyl)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)carbamate(Intermediate B6(P), 532 mg) as a white foam.

LCMS m/z 648 (M+H)⁺ (ES⁺); 646 (M−H)⁻ (ES⁻)

TFA (1.5 mL, 19.47 mmol) was added to a solution of Intermediate B6(P)(530 mg, 0.818 mmol) in DCM (5 mL) and the mixture was stirred at rt for35 minutes. The mixture was diluted with toluene (100 mL) andconcentrated under reduced pressure. The residue was redissolved in DCM(20 mL) and washed with saturated NaHCO₃ solution (20 mL) followed bysaturated brine (20 mL). The organic phase was dried (MgSO₄) andconcentrated under reduced pressure to yield Intermediate B6 (440 mg) asa brown foam.

LCMS m/z 548 (M+H)⁺ (ES⁺); 546 (M−H)⁻ (ES⁻)

Intermediate C11-(4-(2-Chloropyrimidin-4-yloxy)naphthalen-1-yl)-3-(3-isopropyl-1-p-tolyl-1H-pyrazol-5-yl)urea

To a solution of Intermediate G2 (5.00 g, 18.4 mmol) in a mixture ofisopropyl acetate (50 mL) and anhydrous THF (50 mL) was addedportion-wise phenyl(3-isopropyl-1-(p-tolyl)-1H-pyrazol-5-yl)carbamateIntermediate A2* (7.72 g, 23.0 mmol) followed by triethylamine (0.64 mL,4.6 mmol) and the reaction mixture maintained at RT for 18 hr. Duringthis interval a thick purple precipitate formed which was collected byfiltration and then washed with a mixture of isopropyl acetate and THF(1:1 v/v, 3×40 mL). The solid was purified by flash columnchromatography (SiO₂, 330 g, 0-5% MeOH in DCM, gradient elution) toafford the title compound, Intermediate C1 as a pale purple solid (5.72g, 47%); R^(t) 2.48 min (Method 4); m/z 513 (M+H)⁺ (ES⁺).

Intermediate C21-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)urea

A stirred suspension of Intermediate A1* (3 g, 8.59 mmol) andIntermediate G2 (2.333 g, 8.59 mmol) in isopropyl acetate (100 mL) wastreated with triethylamine (0.3 mL, 2.152 mmol) and stirred at 60° C.(bath) for 1 h. The solution was diluted with ethyl acetate (300 mL),washed with water (2×100 mL) followed by brine (100 mL), was dried(Na₂SO₄) and evaporated. The residue was purified on a 220 g redisepsilica cartridge using 5%, for 17 column volumes, and then 40% ofacetone in toluene as eluent and then on another 220 g redisep silicacartridge using 0 to 3% MeOH/DCM as eluent to give Intermediate C2(3.703 g) as a buff foam.

¹H NMR (400 MHz, DMSO-d6) δ 9.14 (s, 1H), 8.79 (s, 1H), 8.65 (d, 1H),8.09 (d, 1H), 7.96 (d, 1H), 7.79 (d, 1H), 7.67-7.64 (m, 1H), 7.60-7.56(m, 1H), 7.47-7.37 (m, 5H), 7.26 (d, 1H), 6.41 (s, 1H), 2.40 (s, 3H),1.28 (s, 9H).

LCMS m/z 527/529 (M+H)⁺ (ES⁺)

Intermediate C31-(3-(tert-Butyl)-1-(4-methoxyphenyl)-1H-pyrazol-5-yl)-3-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)urea

In a 100 mL flask, a solution of Intermediate A3* (1917 mg, 5.24 mmol)and Intermediate G2 (1500 mg, 5.24 mmol) in isopropyl acetate (58 mL)was treated with triethylamine (113 μL, 0.813 mmol). The resultant brownsolution was heated at 70° C. for 2 h then the solvent removed in vacuoto afford a thick brown oil. The crude product was purified bychromatography on silica gel (120 g column, EtOAc 0-15% in DCM) toafford Intermediate C3 (2.169 g) as a white crystalline solid.

¹H NMR (400 MHz, DMSO-d6) δ 9.14 (s, 1H), 8.75 (s, 1H), 8.66 (d, 1H),8.09 (d, 1H), 7.97 (d, 1H), 7.82-7.77 (m, 1H), 7.69-7.62 (m, 1H), 7.58(ddd, 1H), 7.51-7.46 (m, 2H), 7.43 (d, 1H), 7.27 (d, 1H), 7.15-7.10 (m,2H), 6.40 (s, 1H), 3.84 (s, 3H), 1.29 (s, 9H).

LCMS m/z 544 (M+H)⁺ (ES⁺)

Intermediate C4(H)1-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-chloropyrimidin-4-yl)oxy)-5,6,7,8-tetrahydronaphthalen-1-yl)urea

A mixture of Intermediate A1* (760 mg, 2.176 mmol), Intermediate G3 (500mg, 1.813 mmol) and Et₃N (80 μL, 0.574 mmol) in iPrOAc (15 mL) washeated at 60° C. for 1.5 h. The mixture was cooled, evaporated underreduced pressure and the residue purified by chromatography on silicagel (80 g column, 0-50% EtOAc/isohexane) to afford Intermediate C4(H)(807 mg) as a light brown foam.

¹H NMR (CDCl₃) 400 MHz, δ: 8.42 (d, 1H), 7.43 (d, 1H), 7.32 (d, 2H),7.22 (d, 2H), 6.88 (d, 1H), 6.74 (d, 1H), 6.50 (s, 1H), 6.44 (s, 1H),6.37 (s, 1H), 2.48 (t, 2H), 2.41 (t, 2H), 2.36 (s, 3H), 1.73-1.62 (m,4H), 1.35 (s, 9H).

LCMS m/z 531/3 (M+H)⁺ (ES⁺)

Intermediate D13-Amino-5-bromo-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide

Method 1

T3P (1-propanephosphonic acid cyclic anhydride 50 wt % in EtOAc, 4.13ml, 6.94 mmol) was added carefully to a solution of3-amino-5-bromobenzoic acid (1 g, 4.63 mmol),2-(2-(2-methoxyethoxy)ethoxy)ethanamine (1.022 ml, 6.13 mmol) and TEA(1.936 ml, 13.89 mmol) in DCM (20 mL). Ice bath used sporadically toprevent temperature rising above 35° C. Stirred at rt for 1 h thenpartitioned with sat. NaHCO₃ soln. (20 mL). Aqueous layer was separatedand partitioned with fresh DCM (20 mL), organics separated, bulked andpartitioned with 20% w/w NaCl soln. (20 mL). Organic layer wasseparated, dried (MgSO₄), filtered and the solvent evaporated. The crudeproduct was purified by chromatography on the Companion 40 g column, 2%MeOH:DCM to 5%) to afford Intermediate D1 (1.5 g) as a thick, colourlessoil.

¹H NMR (400 MHz, DMSO-d6) δ 8.37 (t, 1H), 7.08 (t, 1H), 7.00 (dd, 1H),6.85 (t, 1H), 5.58 (s, 2H), 3.57-3.46 (m, 8H), 3.45-3.39 (m, 2H),3.39-3.34 (m, 2H), 3.23 (s, 3H).

LCMS m/z 361/363 (M+H)⁺ (ES⁺)

Method 2

3-Bromo-5-nitrobenzoic acid (725 g, 2.947 mol) was refluxed in thionylchloride (2.365 kg, 19.88 mol) until a clear (brown) solution obtained(takes about 3-4 h). [NOTE: HCl is evolved, and so a large NaOH(aq)scrubber is required, with ice/water cooling]. The solution wasconcentrated in vacuo to provide the (brown) acid chloride(3-bromo-5-nitrobenzoyl chloride). The acid chloride was dissolved inethyl acetate (500 mL) and added as a small, controlled stream to2-(2-(2-methoxyethoxy)ethoxy)ethanamine (481 g, 2.947 mol) dissolved inethyl acetate (4 L) and triethylamine (447.31 g, 4.42 mol)) at 15° C.over a period of about 1 h. The resulting heavy suspension was stirredovernight at rt, after which brine (5 L) was added to the reactionmixture and the aqueous phase was separated. The aqueous washings werere-extracted with ethyl acetate and then the combined organics werewashed with water (1×5 L), dried (MgSO4), and passed through a silicaplug (washed with 1×500 mL fresh ethyl acetate). The solvent wasevaporated to provide a deep brown oil, to which was added diethyl ether(2 L). The resulting mixture was cooled in an ice bath and stirred for 1h, after which the solid thereby obtained was washed twice with diethylether and then dried. The product(3-nitro-5-bromo-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-benzamide,Intermediate Dr) was obtained as 850 g (73.3%) of a pale brown,amorphous, free flowing solid that was used in the next step without anyfurther purification.

¹H NMR (400 MHz, DMSO-d6) δ 9.04 (t, 1H), 8.66 (dd, 1H), 8.55 (t, 1H),8.47 (t, 1H), 3.60-3.43 (m, 10H), 3.42-3.37 (m, 2H), 3.21 (s, 3H).

LCMS m/z 391/393 (M+H)⁺ (ES⁺); 389/391 (M−H)⁻ (ES⁻)

Intermediate D1*

(250 g, 0.639 mol) was placed in a 4 L autoclave with Pt/C (5%) and IMS(2 L). The mixture was then heated to 80° C. under H₂ (40 atm) for 48 h.The resulting mixture was then cooled and filtered and the residue waswashed with fresh IMS. The solvent was evaporated from the combinedorganics to provide an orange/red oil. This oil was triturated withdiethyl ether to provide a suspension of a solid that was filtered,washed with further diethyl ether and then dried. This affordedIntermediate D1 (207.8 g, 90%) as an off-white, amorphous solid that had¹H NMR and LCMS data essentially identical to those of the materialobtained via Method 1 above.

Intermediate D23-Amino-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide

Method 1

Pd(PPh₃)₄ (0.240 g, 0.208 mmol) was added to a degassed suspension ofIntermediate D1 (1.5 g, 4.15 mmol), CuI (0.040 g, 0.208 mmol), andethynyltriisopropylsilane (1.397 ml, 6.23 mmol) in TEA (2 mL) and DMF(10 mL). Heated at 80° C. (block temp.) for 4 h then cooled and filtered(Whatman glass fibre pad GF/A). Solvents were evaporated and the residuepartitioned between EtOAc (200 mL) and 20% w/w NaCl soln. (250 mL)Organic layer separated, dried (MgSO₄), filtered and solvent evaporatedto a thick brown oil found to be an impure mixture of starting materialand product. This mixture was subjected to the same reaction conditionsand work up as before for 1 h. The crude product was purified bychromatography on the Companion 40 g column, MeOH:EtOAc to 0% to 5%) toafford3-amino-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-5-((triisopropylsilyl)ethynyl)benzamide(1.5 g) as a thick, yellow oil.

¹H NMR (400 MHz, DMSO-d6) δ 8.41 (t, 1H), 7.10-7.01 (m, 2H), 6.79 (dd,1H), 5.45 (s, 2H), 3.55-3.47 (m, 8H), 3.44-3.35 (m, 4H), 3.22 (s, 3H),1.10 (s, 21H).

LCMS m/z 463 (M+H)⁺ (ES⁺)

The (triisopropylsilyl)ethynyl-substituted benzamide obtainedimmediately above (1.5 g, 3.24 mmol) was dissolved in EtOAc (15 mL) andTBAF, 1M in THF (3.24 ml, 3.24 mmol) added. Stirred for 1 h thenpartitioned between water (10 mL) and ethyl acetate (10 mL). The organiclayer separated and washed with 20% w/w NaCl soln. (20 mL), dried(MgSO₄), filtered and evaporated. The crude product was purified bychromatography on silica gel (40 g column, 1% MeOH:DCM to 6%) to affordIntermediate D2 (750 mg) as a clear, yellow oil.

¹H NMR (400 MHz, DMSO-d6) δ 8.36 (t, 1H), 7.12-7.02 (m, 2H), 6.76 (dd,1H), 5.45 (s, 2H), 4.07 (s, 1H), 3.58-3.46 (m, 8H), 3.45-3.36 (m, 4H),3.23 (s, 3H).

LCMS m/z 307 (M+H)⁺ (ES⁺)

Method 2

Pd(PPh₃)₄ (90 g, 78 mmol) was added to a degassed (N₂ purging for 30mins) solution of Intermediate D1 (500 g, 1.384 mol), CuI (13.7 g, 72mmol), ethynyltriisopropylsilane (227.68 g, 1.523 mol) and TEA (667.92g, 6.601 mol) in DMF (4 L). The mixture was heated to 85° C. under N₂for 7 h before being allowed to cool overnight. As much DMF solvent aspossible was removed in vacuo to provide a residue that was then takenup in ethyl acetate. The resulting solution was passed through largesilica plug to remove inorganic impurities. The silica plug was washedtwice with ethyl acetate. The combined organics were stirred with 1 L ofconc. HCl/water (50%) for 10 mins. The aqueous layer was separated andthe ethyl acetate layer was discarded (after checking by TLC for anyremaining product). The acidic aqueous layer was back washed withdiethyl ether (×2), and the organic washings were discarded. The aqueouslayer was then basified cautiously with NaOH until the pH reachedapproximately 9 to 10. The aqueous layer was then extracted twice withethyl acetate. The combined organics were washed with water, dried(MgSO₄) and passed through a silica plug before the solvent wasevaporated to provide3-amino-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-5-((triisopropylsilyl)-ethynyl)benzamide(489.4 g, 76.4%) as a viscous orange/red oil that had ¹H NMR and LCMSdata essentially identical to those of the material obtained via Method1 above, and which oil was used in the next step without furtherpurification.

The (triisopropylsilyl)ethynyl-substituted benzamide obtainedimmediately above (290 g, 627 mmol, 1 eq.) was dissolved in EtOAc (2.5L) and TBAF, 1 M in THF (690.7 mL, 689 mmol, 1.1 eq.) was added in oneportion. The resulting mixture was stirred overnight at rt before thesolvent was evaporated. The residue was dissolved in fresh ethyl acetatebefore being washed with a solution of sodium hexafluorophosphate (210g, 2 eq.) in water (750 mL). The aqueous and organic layers were thenseparated. The organic layer was dried (MgSO₄) and passed through asilica plug. The solvent was then evaporated to provide a solid that wasfound to contain some TBAF. The solid was then slurried in a smallquantity of ethyl acetate and passed through a further silica plug thatwas washed sparingly with ethyl acetate. Solvent removed from thefiltrate in vacuo to provide Intermediate D2 (155.76 g, 81.1%) as aviscous, orange/red oil of about 90% purity that had ¹H NMR and LCMSdata essentially identical to those of the material obtained via Method1 above, and which oil was used without further purification.

Intermediate D33-Amino-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-benzamide

3-Amino-5-methoxybenzoic acid (1.0 g, 5.98 mmol) was added to an icecold suspension of 2-(2-(2-methoxyethoxy)ethoxy)ethanamine (1.2 g, 7.35mmol), 50% T3P in ethyl acetate (4.50 ml, 7.56 mmol) and TEA (2.5 ml,17.94 mmol) in ethyl acetate (15 mL). The mixture was allowed to warm tort and stir overnight. Saturated NaHCO₃ (20 mL) was added and themixture was extracted with ethyl acetate (3×10 mL). The combined organicphases were washed with saturated brine (20 mL), dried (MgSO₄) andconcentrated under reduced pressure to yield a yellow oil. The oil waspurified by chromatography on the Companion (40 g column, 0-100%acetone/toluene) to afford a pale yellow oil. The oil was purified bychromatography on the Companion (40 g column, 0-100% THF/DCM) to affordthe subtitle compound (843 mg) as a pale yellow oil.

LCMS m/z 313 (M+H)⁺ (ES⁺)

Intermediate D4 3-Amino-5-bromo-N-(2-(2-methoxyethoxy)ethyl)benzamide

A stirred mixture of 3-amino-5-bromobenzoic acid (800 mg, 3.59 mmol),2-(2-methoxy-ethoxy)ethanamine (856 mg, 7.18 mmol) and triethylamine(1.5 mL, 10.76 mmol) in DCM (13 mL) was cooled in an ice bath. 50 wt %T3P in EtOAc (3.2 mL, 5.38 mmol) was added dropwise, the ice bath wasremoved and the reaction mixture allowed to warm to rt. DMF (2 mL) wasadded to aid solubility and the reaction stirred at rt overnight. Thereaction mixture was partitioned between sat. aq. NaHCO₃ (50 mL) and DCM(50 mL). The aqueous phase was back extracted with fresh DCM (50 mL).The combined organic extracts were washed with water (100 mL), brine(100 mL), dried (MgSO₄), filtered and concentrated in vacuo to afford anorange oil. The crude product was purified by chromatography on silicagel (40 g column, 0-5% MeOH) to afford Intermediate D4 (880 mg) as anorange oil.

¹H NMR (DMSO-d6) 400 MHz, δ: 8.36 (t, 1H), 7.07 (t, 1H), 7.00-6.99 (m,1H), 6.84 (t, 1H), 5.57 (s, 2H), 3.53-3.48 (m, 4H), 3.44-3.42 (m, 2H),3.35 (q, 2H), 3.23 (s, 3H).

LCMS m/z 317/319 (M+H)⁺ (ES⁺)

Intermediate D5 3-Amino-5-ethynyl-N-(2-(2-methoxyethoxy)ethyl)benzamide

To a degassed solution of Intermediate D4 (830 mg, 2.59 mmol),ethynyltriisopropylsilane (880 μL, 3.92 mmol), copper(I) iodide (24.67mg, 0.130 mmol) and TEA (1.55 mL, 11.12 mmol) in DMF (8 mL) was addedPd(PPh₃)₄ (150 mg, 0.130 mmol). The reaction was heated at 85° C. for 3h. The reaction was cooled to rt then partitioned between EtOAc (50 mL)and brine (50 mL). The aqueous phase was back extracted with EtOAc (50mL). The combined organic extracts were washed with brine (100 mL),dried (MgSO₄), filtered and concentrated in vacuo to afford a brownsemi-solid (1.65 g). The crude product was purified by chromatography onsilica gel (80 g column, 0-3% MeOH in DCM) to afford3-amino-N-(2-(2-methoxyethoxy)ethyl)-5-((triisopropylsilyl)ethynyl)benzamide(796 mg) as a beige solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 8.39 (t, 1H), 7.05-7.04 (m, 1H), 7.03 (t,1H), 6.79-6.78 (m, 1H), 5.44 (br s, 2H), 3.53-3.48 (m, 4H), 3.44-3.42(m, 2H), 3.35 (q, 2H), 3.23 (s, 3H), 1.10 (s, 21H).

LCMS m/z 419 (M+H)⁺ (ES⁺)

To a stirred solution of the (triisopropylsilyl)ethynyl-substitutedbenzamide obtained immediately above (717 mg, 1.473 mmol) in EtOAc (9mL) was added 1M TBAF in THF (1473 μL, 1.473 mmol). The reaction wasstirred at rt for 1 h. The reaction mixture was partitioned betweenwater (30 mL) and EtOAc (20 mL). The organic layer was washed with brine(20 mL), dried (MgSO₄), filtered and concentrated to afford a brown oil.The crude product was dissolved in the minimum quantity of MeOH andloaded onto SCX. The column was eluted with MeOH followed by 1% NH₃ inMeOH. The filtrate was concentrated in vacuo to afford a brown oil at˜70% purity. The crude product was purified by chromatography on silicagel (40 g column, 0-5% MeOH in DCM) to afford Intermediate D5 (377 mg)as an orange oil.

¹H NMR (DMSO-d6) 400 MHz, δ: 8.36 (t, 1H), 7.06-7.04 (m, 2H), 6.75-6.74(m, 1H), 5.45 (s, 2H), 4.07 (s, 1H), 3.53-3.47 (m, 4H), 3.44-3.42 (m,2H), 3.37-3.33 (m, 2H), 3.23 (s, 3H).

LCMS m/z 263 (M+H)⁺ (ES⁺)

Intermediate G1(P)tert-Butyl(4((2-chloropyridin-4-yl)oxy)naphthalen-1-yl)carbamate

A mixture of Intermediate G1 (1000 mg, 3.69 mmol) di-tert-butyldicarbonate (750 mg, 3.44 mmol) in t-BuOH (10 mL) was stirred at refluxfor 18 h. The mixture was diluted with water (15 mL) and collected byfiltration. The solid was triturated in diethyl ether to yieldIntermediate G1(P)(1002 mg) as a pale grey solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.37 (s, 1H), 8.28 (d, 1H), 8.16 (d, 1H),8.82 (dd, 1H), 7.66 (d, 1H), 7.66-7.54 (m, 2H), 7.40 (d, 1H), 7.03 (d,1H), 6.91 (dd, 1H), 1.52 (s, 9H). LCMS m/z 371 (M+H)⁺ (ES⁺); 369 (M−H)⁻(ES⁻)

Intermediate G34-((2-Chloropyrimidin-4-yl)oxy)-5,6,7,8-tetrahydronaphthalen-1-amine

2,4-Dichloropyrimidine (0.958 g, 6.43 mmol) was added to a stirredmixture of 4-amino-5,6,7,8-tetrahydronaphthalen-1-ol (1 g, 6.13 mmol)and DBU (1.1 mL, 7.30 mmol) at 0-5° C. The mixture was stirred for 3 hthen more DBU (0.6 mL) and 2,4-dichloropyrimidine (300 mg) added andstirred for a further 2 h. The mixture was partitioned between EtOAc(100 mL) and water (50 mL), the organic layer was washed with water (50mL), dried (MgSO₄), filtered and evaporated under reduced pressure. Thecrude product was purified by chromatography on silica gel (80 g column,0-50% EtOAc/isohexane) to afford Intermediate G3 (510 mg) as brownsolid.

¹H NMR (CDCl₃) 400 MHz, δ: 8.36 (d, 1H), 6.74 (d, 1H), 6.62 (d, 1H),6.56 (d, 1H), 3.60 (s, 2H), 2.49-2.46 (m, 4H), 1.87-1.81 (m, 2H),1.74-1.68 (m, 2H).

LCMS m/z 276/8 (M+H)⁺ (ES⁺)

Intermediate J1(P)3-((4-((4-((tert-Butoxycarbonyl)amino)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynylbenzoicacid

A suspension of Intermediate G2(P) (42.6 g, 115 mmol), Intermediate K1*(40.00 g, 126 mmol), BINAP (6.42 g, 10.31 mmol) and caesium carbonate(74.6 g, 229 mmol) in 1,4-dioxane (500 mL) was degassed with nitrogenfor 10 minutes. Pd₂(dba)₃ (4.20 g, 4.58 mmol) was added and the mixturewas heated to 90° C. for 2.5 h. The mixture was diluted with diethylether (600 mL) then washed with water (600 mL), followed by 0.5 M HClsolution (500 mL) and saturated brine (500 mL). The organic phase wasdried (MgSO₄), filtered and concentrated in vacuo affording3-((4-((4-((tert-butoxycarbonyl)amino)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-((triisopropylsilyl)ethynyl)benzoicacid (Intermediate J1(P)*, 96 g) as a red foam which was used withoutfurther purification.

Intermediate J1(P)* (96 g) was dissolved in THF (60 mL) and diluted withMeCN (400 mL). 1.0 M TBAF in THF (235 mL, 235 mmol) was added and thereaction stirred at rt overnight. The reaction was diluted with MeCN(300 mL) and water (600 mL), then 1M HCl solution (100 mL, 1 eq.) wasadded and stirring continued resulting in the precipitation of a pinksolid which was collected by filtration. The pink solid was trituratedin MeCN at 80° C., collected by filtration and dried at 40° C. undervacuum for 2 h. The solid was re-suspended in (9:1) EtOAc/THF (400 ml)and heated to 60° C. for 90 mins then cooled to room temperature andstirred overnight. The suspended solid was collected by filtration,washing with EtOAc affording Intermediate J1(P) (47 g) as a paleyellow/beige solid.

¹H NMR (400 MHz, DMSO-d6) δ: 13.12 (bs, 1H), 9.83 (s, 1H), 9.32 (s, 1H),8.46 (d, 1H), 8.28 (s, 1H), 8.10 (d, 1H), 8.01 (s, 1H), 7.82 (d, 1H),7.54-7.63 (m, 3H), 7.49 (s, 1H), 7.42 (d, 1H), 6.61 (d, 1H), 4.17 (s,1H), 1.52 (s, 9H).

LCMS m/z 497 (M+H)⁺ (ES⁺); 495 (M−H)⁻ (ES⁻)

Intermediate J2(P)3-((4-((4-((tert-Butoxycarbonyl)amino)naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-ethynylbenzoicacid

N₂ was bubbled through a mixture of Intermediate G1(P) (0.5 g, 1.348mmol), 3-amino-5-((triisopropylsilyl)ethynyl)benzoic acid (0.490 g,1.544 mmol), Cs₂CO₃ (0.966 g, 2.97 mmol), BINAP (0.078 g, 0.125 mmol)and Pd₂dba₃ (0.056 g, 0.061 mmol) in dioxane (15 mL) for 10 min thenheated at 90C for 4 h. The mixture was partitioned between ether (100mL) and 1M HCl (50 mL), the organic layer separated, washed with water,dried (MgSO₄), filtered and evaporated under reduced pressure. Theresidue was triturated with ether/isohexane, filtered and dried toafford3-((4-((4-((tert-butoxycarbonyl)amino)naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-((triisopropylsilyl)ethynyl)benzoicacid (760 mg) which was used crude in the next step.

1.0 M TBAF in THF (2.5 ml, 2.500 mmol) was added to a stirred solutionof3-((4-((4-((tert-butoxycarbonyl)amino)naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-((triisopropylsilyl)ethynyl)-benzoicacid obtained immediately above (760 mg) in THF (15 mL). The mixture wasstirred for 2 h then water (10 mL) added and acidified to pH-4 with 1MHCl. The mixture was partitioned between EtOAc (70 mL) and water (40mL), the organic phase washed with sat brine (50 mL), dried (MgSO₄),filtered and evaporated under reduced pressure. The crude product waspurified by chromatography on silica gel (40 g column, 0-100%EtOAc/isohexane) to afford Intermediate J2(P) (344 mg) as a foam.

¹H NMR (DMSO-d6) 400 MHz, δ: 13.07 (s, 1H), 9.39 (s, 1H), 9.29 (s, 1H),8.18-8.13 (m, 4H), 7.84 (d, 1H), 7.66-7.56 (m, 3H), 7.44 (s, 1H), 7.38(d, 1H), 6.66 (dd, 1H), 6.07 (d, 1H), 4.22 (s, 1H), 1.53 (s, 9H).

LCMS m/z 496 (M+H)⁺ (ES⁺)

Intermediate J3(P)3-((4-((4-((tert-Butoxycarbonyl)amino)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-methoxybenzoicacid

N₂ was bubbled through a stirred mixture of Intermediate G2(P) (10 g,26.9 mmol), 3-amino-5-methoxybenzoic acid (8.99 g, 53.8 mmol) and p-TSAmonohydrate (1.02 g, 5.36 mmol) in THF (150 mL) for 10 min. The mixturewas heated under reflux for 20 h, cooled and filtered. The filtrate wasevaporated, MeOH (300 mL) added and the solid filtered, washed with MeOHthen ether to afford the sub-title compound (10.063 g).

¹H NMR (400 MHz; DMSO-d6) δ 12.83 (brs, 1H), 9.68 (s, 1H), 9.32 (s, 1H),8.44 (d, 1H), 8.11 (d, 1H), 8.13-8.10 (m, 2H), 7.61-7.51 (m, 4H), 7.41(d, 1H), 6.98 (s, 1H), 6.58 (d, 1H), 3.60 (s, 3H), 1.52 (s, 9H).

LCMS m/z 503 (M+H)⁺ (ES⁺)

Intermediate K1* 3-Amino-5-((triisopropylsilyl)ethynyl)benzoic acid

Pd(PPh₃)₄ (9.36 g, 8.10 mmol) was added to a degassed suspension of3-amino-5-bromobenzoic acid (50 g, 231 mmol), CuI (1.499 g, 7.87 mmol),and ethynyltriisopropylsilane (80 mL, 356 mmol) in Et₃N (300 mL) and DMF(300 mL). The mixture was heated to 90° C. for 2 h. The mixture wascooled and carefully poured into ice-cold HCl (2.0 M aq.; 1100 mL, 2200mmol) and diluted with diethyl ether (500 mL). The biphasic mixture wasfiltered to remove palladium residues. The layers of the filtrate wereseparated and the aqueous phase was extracted with a further portion ofdiethyl ether (300 mL). The organic phases were combined and washed with20% brine (2×300 mL), 40% brine (300 mL), dried (MgSO₄), filtered andconcentrated in vacuo affording a pale orange solid. The solid wasrecrystallised in acetonitrile (250 mL) and collected by filtration,washing with fresh acetonitrile (2×30 mL) affording the product as ayellow solid. The solid was slurried in hexane (250 mL) for 5 h thenfiltered, washing with more hexane to afford Intermediate K1* (45.5 g)as a pale yellow solid.

¹H NMR (400 MHz, DMSO-d6) δ: 12.87 (bs, 1H), 7.18 (t, 1H), 7.10 (t, 1H),6.86 (t, 1H), 5.54 (bs, 2H), 1.10 (s, 21H).

LCMS m/z 318 (M+H)⁺ (ES⁺); 316 (M−H)⁻ (ES⁻)

Intermediate M13-(tert-Butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazole-5-carboxylic acid

Pyridine (350 μL, 4.33 mmol) followed by activated 4A molecular sieves(0.5 g) were added to a stirred mixture of(4-(dimethylamino)phenyl)boronic acid (575 mg, 3.48 mmol), ethyl3-(tert-butyl)-1H-pyrazole-5-carboxylate (425 mg, 2.166 mmol) and copper(II) acetate (590 mg, 3.25 mmol) in DCM (15 mL) at rt. open to the air.The mixture was stirred for 4 h. A mixture of ether/isohexane (3:1, 300mL) was added and the solid was filtered off. The filtrate wasevaporated under reduced pressure and the residue was purified bychromatography on the Companion (80 g column, 0-60% ether/isohexane) toafford ethyl3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazole-5-carboxylate(Intermediate M(P)1; 464 mg) as a colourless oil.

LCMS m/z 316 (M+H)⁺ (ES⁺)

1 M sodium hydroxide solution (1.5 ml, 1.500 mmol) was added to astirred solution of Intermediate M(P)1 (0.46 g, 1.458 mmol) in THF (3mL) at rt. The mixture was stirred for 3 h at rt then methanol (1 mL)was added and the mixture was stirred for a further 1 h. The mixture wasthen heated to 40° C. for 1 h, diluted with water (10 mL) and washedwith diethyl ether (2×10 mL). The aqueous phase was treated with 1 M HCl(1.5 mL) and extracted with ethyl acetate (3×10 mL). The combinedorganic phases were washed with saturated brine (10 mL), dried (MgSO₄)and concentrated to yield Intermediate M1 (395 mg) as an off-whitesolid.

¹H NMR (400 MHz; CDCl₃) δ: 7.28-7.22 (m, 2H), 6.91 (s, 1H), 6.74-6.67(m, 2H), 2.98 (s, 6H), 1.35 (s, 9H).

LCMS m/z 288 (M+H)⁺ (ES⁺); 286 (M−H)⁻ (ES⁻)

Intermediate M23-(tert-Butyl)-1-(2,4-dimethoxyphenyl)-1H-pyrazole-5-carboxylic acid

Pyridine (2.3 mL, 28.4 mmol) followed by activated 4A molecular sieves(2 g) were added to a stirred mixture of (2,4-dimethoxyphenyl)boronicacid (3.88 g, 21.31 mmol), ethyl3-(tert-butyl)-1H-pyrazole-5-carboxylate (2.79 g, 14.20 mmol) and copper(II) acetate (3.87 g, 21.31 mmol) in DCM (50 mL) at rt. open to the air.The mixture was stirred for 3 days then a mixture of ether/isohexane(3:1, 300 mL) was added and the solid was filtered off. The filtrate wasevaporated under reduced pressure and the residue was purified bychromatography on silica gel (120 g column, 0-20% EtOAc/isohexane) toafford ethyl3-(tert-butyl)-1-(2,4-dimethoxyphenyl)-1H-pyrazole-5-carboxylate(Intermediate M(P)2; 377 mg) as an oil.

¹H NMR (CDCl₃) 400 MHz, δ: 7.31 (d, 1H), 6.84 (s, 1H), 6.56 (dd, 1H),6.54 (d, 1H), 4.21 (q, 2H), 3.86 (s, 3H), 3.73 (s, 3H), 1.38 (s, 9H),1.24 (t, 3H).

LCMS m/z 333 (M+H)⁺ (ES⁺)

A mixture of Intermediate M(P)2 (365 mg, 0.933 mmol), LiOH (70 mg, 2.92mmol) in THF (5 mL) and water (2 mL) was stirred at rt for 4 h. EtOH (5mL) was added and the mixture stirred at rt for 18 h. The solvent wasevaporated and the residue partitioned between EtOAc (30 mL) and aq 1MHCl (30 mL). The organic layer was washed with water (10 mL), dried(MgSO₄), filtered and evaporated under reduced pressure to give a gumthat was triturated with ether/isohexane to afford Intermediate M2 (220mg) as a white solid.

¹H NMR (CDCl₃) 400 MHz, δ: 7.32 (d, 1H), 6.91 (s, 1H), 6.56 (dd, 1H),6.52 (d, 1H), 3.86 (s, 3H), 3.72 (s, 3H), 1.38 (s, 9H).

LCMS m/z 305 (M+H)⁺ (ES⁺); 303 (M−H)⁻ (ES⁻)

Intermediate M(P)3 Ethyl3-(tert-butyl)-1-(3-(hydroxymethyl)phenyl)-1H-pyrazole-5-carboxylate

To a stirred mixture of ethyl 3-(tert-butyl)-1H-pyrazole-5-carboxylate(4.39 g, 22.37 mmol), (3-(hydroxymethyl)phenyl)boronic acid (5.1 g, 33.6mmol) and copper (II) acetate (6.10 g, 33.6 mmol) in DCM (130 mL) wasadded pyridine (3.62 mL, 44.7 mmol) followed by 4A molecular sieves. Theresulting mixture was stirred at rt open to the air for 48 h. Thereaction mixture was filtered and the filtrate concentrated in vacuo.Et₂O (200 mL) was added, the resulting mixture filtered and thefiltrated concentrated in vacuo to afford a green oil. The crude productwas purified by chromatography on silica gel (120 g column, 0-40% EtOAcin isohexane) to afford Intermediate M(P)3 (6.2 g) as an oil.

¹H NMR (DMSO-d6) 400 MHz, δ: 7.42-7.35 (m, 3H), 7.27-7.24 (m, 1H), 6.97(s, 1H), 5.31 (t, 1H), 4.56 (d, 2H), 4.17 (q, 2H), 1.30 (s, 9H), 1.16(t, 3H).

LCMS m/z 303 (M+H)⁺ (ES⁺)

Intermediate M(P)4 Ethyl3-(tert-butyl)-1-(3-(chloromethyl)phenyl)-1H-pyrazole-5-carboxylate

To a stirred solution of Intermediate M(P)3 (6.2 g, 20.50 mmol) in DCM(10 mL) was added SOCl₂ (1 M in DCM, 41.0 mL, 41.0 mmol). The resultingsolution was stirred at rt for 2 h. The solvent was removed in vacuo andthe crude product was purified by chromatography on silica gel (220 gcolumn, 0-100% EtOAc in isohexane) to afford Intermediate M(P)4 (5.64 g)as an oil.

¹H NMR (DMSO-d6) 400 MHz, δ: 7.52-7.45 (m, 3H) 7.41-7.38 (m, 1H) 7.00(s, 1H) 4.83 (s, 2H) 4.17 (q, 2H) 1.30 (s, 9H) 1.16 (t, 3H).

LCMS m/z 321 (M+H)⁺ (ES⁺)

Intermediate M53-(tert-Butyl)-1-(3-((dimethylphosphoryl)methyl)phenyl)-1H-pyrazole-5-carboxylicacid

A suspension of Intermediate M(P)4 (3.65 g, 10.81 mmol), xantphos (0.375g, 0.649 mmol), palladium(II) acetate (0.121 g, 0.540 mmol), potassiumphosphate, tribasic (2.52 g, 11.89 mmol) and dimethylphosphine oxide(0.928 g, 11.89 mmol) in DMF (30 mL) was purged with N₂ for 20 mins. Thereaction mixture was heated at 110° C. for 1 h, cooled to rt thenpartitioned between DCM (300 mL) and water (200 mL). The organic layerwas washed with water (2×200 mL), brine (300 mL), dried (MgSO₄),filtered and concentrated in vacuo. The crude product was purified bychromatography on silica gel (120 g column, 0-10% MeOH/DCM) to affordethyl3-(tert-butyl)-1-(3-((dimethylphosphoryl)methyl)phenyl)-1H-pyrazole-5-carboxylate(Intermediate M(P)5, 2.29 g) as an oil which solidified on standing.

¹H NMR (CDCl₃) 400 MHz, δ: 7.43-7.27 (m, 4H), 6.87 (s, 1H), 4.22 (q,2H), 3.23 (d, 2H), 1.48 (d, 6H), 1.36 (s, 9H), 1.28 (t, 3H).

LCMS m/z 363 (M+H)⁺ (ES⁺)

Sodium hydroxide (1 M aq.) (12.0 mL, 12.00 mmol) was added to a stirredsolution of Intermediate M(P)5 (2.2 g, 6.07 mmol) in EtOH (40 mL). Theresulting mixture was stirred at rt for 3 h. The solvent was removed invacuo and the resulting residue partitioned between 1M HCl (90 mL) andEtOAc (300 mL). The organic layer was washed with brine (150 mL), dried(MgSO₄), filtered and concentrated in vacuo to afford Intermediate M5(1.61 g) as a cream solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 13.18 (bs, 1H), 7.39-7.43 (m, 1H),7.29-7.31 (m, 3H), 6.93 (s, 1H), 3.23 (d, 2H), 1.36 (d, 6H), 1.30 (s,9H).

LCMS m/z 335 (M+H)⁺ (ES⁺); 333 (M−H)⁻ (ES⁻)

Compound Examples of the Invention Example 13-Ethynyl-5-((4-((4-(3-(3-isopropyl-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-benzamide

Intermediate C1 (146 mg, 0.285 mmol) was dissolved in DMF (3 mL) andadded to Intermediate D2 (87 mg, 0.285 mmol) and p-TSA monohydrate (27.1mg, 0.142 mmol). Stirred at 70° C. (block temperature) for 7 h thenpoured into sat. NaHCO₃ solution (20 mL) and the product extracted withEtOAc (2×20 mL). Organics bulked and washed with 20% w/w brine solution(20 mL), dried (MgSO₄), filtered and evaporated to a yellow solid.

The crude product was preabsorbed onto silica (4 g) and purified bychromatography on silica gel (40 g column, 1% MeOH:DCM to 6%) to afforda pale brown solid. Triturated 4 times with MeCN (2 mL) to afford thetitle compound (60 mg) ¹H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.09(s, 1H), 8.78 (s, 1H), 8.51-8.40 (m, 2H), 8.13-8.01 (m, 2H), 7.94 (d,1H), 7.87 (s, 1H), 7.82 (d, 1H), 7.68-7.54 (m, 2H), 7.53-7.32 (m, 6H),6.56 (d, 1H), 6.38 (s, 1H), 4.11 (s, 1H), 3.58-3.46 (m, 8H), 3.43-3.35(m, 4H), 3.21 (s, 3H), 2.90 (hept, 1H), 2.41 (s, 3H), 1.25 (d, 6H).

LCMS m/z 783 (M+H)⁺ (ES⁺)

Example 23-((4-((4-(3-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-benzamide

Method 1

A suspension of Intermediate C2 (165 mg, 0.282 mmol), Intermediate D2(173 mg, 0.564 mmol) and p-TSA monohydrate (11.0 mg, 0.058 mmol) inTHF/DMF (6 mL, 1:2) was heated at 60° C. overnight. The reaction wascooled to rt and partitioned between EtOAc (40 mL) and sat. aq. NaHCO₃(30 mL). The aqueous layer was extracted with EtOAc (2×40 mL). Thecombined organic extracts were washed with water (2×50 mL), brine (2×50mL), dried (MgSO₄), filtered and concentrated in vacuo. The crudeproduct was purified by chromatography on silica gel (40 g column, 0-10%MeOH) to afford a pale yellow solid. The solid was triturated with EtOAcaffording the title compound (68 mg) as a white solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.75 (s, 1H), 9.08 (s, 1H), 8.76 (s, 1H),8.43-8.46 (m, 2H), 8.06-8.08 (m, 2H), 7.94 (d, 1H), 7.87 (s, 1H), 7.83(d, 1H), 7.56-7.65 (m, 6H), 6.56 (d, 1H), 6.42 (s, 1H), 4.10 (s, 1H),3.48-3.53 (m, 8H), 3.36-3.41 (m, 4H), 3.21 (s, 3H), 2.41 (s, 3H), 1.30(s, 9H).

LCMS m/z 399 (M+2H)²⁺ (ES⁺)

Method 2

TEA (13.38 g, 132 mmol) was added to solution of Intermediate A1* (219g, 627 mmol) and Intermediate B2 (503.11 g, 929 mmol) in isopropylacetate (4 L) in a 10 L jacketed vessel. The resulting mixture washeated to 50-60° C. under stirring. After about 5 minutes, a heavyprecipitate started to form. Stirring was continued for a further 2 h.Analysis by TLC (ethyl acetate) indicated consumption of the startingmaterials. Heating was ceased and the suspension allowed to cool slowlyovernight. The reaction mixture was then filtered through a cloth on alarge diameter (40 cm), large pore clay filter. The solid product waswashed with ethyl acetate and then n-hexane to provide the titlecompound (256 g, 51.2%) as a colourless, amorphous solid that had ¹H NMRand LCMS data essentially identical to those of the material obtainedvia Method 1 above.

Example 33-((4-((4-(3-(3-(tert-Butyl)-1-(4-methoxyphenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide

To a stirred solution of Intermediate C3 (175 mg, 0.316 mmol) andIntermediate D2 (153 mg, 0.474 mmol) in DMF (4 mL) was added p-TSAmonohydrate (30 mg, 0.158 mmol). The resulting solution was heated at60° C. overnight. The reaction was cooled to rt and partitioned betweenEtOAc (30 mL) and sat aq. NaHCO₃ (30 mL). The aqueous phase was backextracted with EtOAc (30 mL). The combined organic extracts were washedwith water (2×50 mL), brine (50 mL), dried (MgSO₄), filtered andconcentrated in vacuo to afford an orange oil (276 mg) at 85% purity.The crude product was purified by chromatography on silica gel (40 gcolumn, 0-10% MeOH in DCM) to afford a pink solid (188 mg), which wastriturated with MeCN to afford the title compound (98 mg) as anoff-white solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.75 (s, 1H), 9.08 (s, 1H), 8.72 (s, 1H),8.46 (t, 1H), 8.43 (d, 1H), 8.06-8.04 (m, 2H), 7.93 (d, 1H), 7.86 (br s,1H), 7.82-7.80 (m, 1H), 7.64-7.54 (m, 2H), 7.50-7.46 (m, 2H), 7.45-7.42(m, 2H), 7.14-7.10 (m, 2H), 6.55 (d, 1H), 6.39 (s, 1H), 4.11 (s, 1H),3.84 (s, 3H), 3.53-3.46 (m, 8H), 3.40-3.35 (m, 4H), 3.20 (s, 3H), 1.28(s, 9H).

LCMS m/z 813 (M+H)⁺ (ES⁺)

Example 43-((4-((4-(3-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-benzamide

Triethylamine (6 μL, 0.043 mmol) was added to a mixture of IntermediateA1* (75 mg, 0.215 mmol) and Intermediate B1 (128 mg, 0.234 mmol) inisopropyl acetate (2 mL) and the mixture heated at 50° C. for 2 h. Thereaction mixture was concentrated under reduced pressure then purifiedby chromatography on the Companion (80 g column, 0-50% acetone/EtOAc) toafford a colourless gum. The gum was purified by chromatography on theCompanion (40 g column, 5% MeOH/DCM) to afford a tan foam. The crudeproduct was purified by preparative HPLC (Waters, Acidic (0.1% Formicacid), Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 25-70% MeCN inWater) to afford the title compound (55 mg) as a white solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.19 (s, 1H), 9.06 (s, 1H), 8.84 (s, 1H),8.34 (dd, 1H), 8.13-8.07 (m, 2H), 7.97 (d, 1H), 7.85 (dd, 1H), 7.69-7.62(m, 1H), 7.62-7.55 (m, 2H), 7.52-7.44 (m, 3H), 7.41-7.33 (m, 3H),6.91-6.86 (m, 1H), 6.55 (dd, 1H), 6.42 (s, 1H), 6.13 (d, 1H), 3.75 (s,3H), 3.56-3.46 (m, 8H), 3.43-3.34 (m, 4H), 3.21 (s, 3H), 2.41 (s, 3H),1.30 (s, 9H).

LCMS m/z 802 (M+H)⁺ (ES⁺); 800 (M−H)⁻ (ES⁻)

Example 53-((4-((4-(3-(3-(tert-Butyl)-1-(4-methoxyphenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-methoxyethoxy)ethyl)-benzamide

To a stirred solution of Intermediate C3 (152 mg, 0.274 mmol) andIntermediate D5 (124 mg, 0.411 mmol) in DMF (4 mL) was added p-TSAmonohydrate (26 mg, 0.137 mmol). The resulting solution was heated at60° C. overnight. The reaction was cooled to rt and partitioned betweenEtOAc (30 mL) and sat aq. NaHCO₃ (30 mL). The aqueous phase was backextracted with EtOAc (30 mL). The combined organic extracts were washedwith water (2×50 mL), brine (50 mL), dried (MgSO₄), filtered andconcentrated in vacuo to afford a dark orange glass. The crude productwas purified by chromatography on silica gel (40 g column, 0-10% MeOH inDCM) to afford a pale pink solid, which was triturated with Et₂O toafford a pale, pink solid (25 mg). The crude product was purified bypreparative HPLC (Waters, Acidic (0.1% Formic acid), Waters X-SelectPrep-C18, 5 μm, 19×50 mm column, 45-65% MeCN in Water) to afford thetitle compound (13 mg) as an off-white solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.77 (s, 1H), 9.14 (s, 1H), 8.79 (s, 1H),8.48 (t, 1H), 8.44 (d, 1H), 8.07-8.04 (m, 2H), 7.93 (d, 1H), 7.86 (br s,1H), 7.81 (d, 1H), 7.64-7.54 (m, 2H), 7.50-7.41 (m, 4H), 7.12 (d, 2H),6.56 (d, 1H), 6.39 (s, 1H), 4.12 (s, 1H), 3.83 (s, 3H), 3.53-3.48 (m,4H), 3.43-3.41 (m, 2H), 2H under water peak at 3.35 ppm, 3.22 (s, 3H),1.28 (s, 9H).

LCMS m/z 385 (M+2H)²⁺ (ES⁺)

Example 63-((4-((4-(3-(3-(tert-Butyl)-1-(2,3,5,6-tetradeutero-4-(trideuteromethyl)-phenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide

A mixture of Intermediate A4* (0.5 g, 1.403 mmol), Intermediate B2(0.760 g, 1.403 mmol) and Et₃N (40 μL, 0.287 mmol) in iPrOAc (20 mL)were stirred at 50° C. for 3 h. The mixture was cooled, filtered and thesolid washed with iPrOAc (15 mL), EtOAc (15 mL) then MeCN (15 mL). Thesolid was dried at 50° C. under vacuum to afford the title compound (745mg) as a white solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.75 (s, 1H), 9.09 (s, 1H), 8.76 (s, 1H),8.47-8.43 (m, 2H), 8.08-8.06 (m, 2H), 7.94 (d, 1H), 7.87 (s, 1H), 7.83(d, 1H), 7.65-7.56 (m, 2H), 7.45 (s, 1H), 7.43 (d, 1H), 6.56 (d, 1H),6.42 (s, 1H), 4.10 (s, 1H), 3.54-3.48 (m, 8H), 3.41-3.36 (m, 4H), 3.21(s, 3H), 1.30 (s, 9H)

LCMS m/z 804 (M+H)⁺ (ES⁺)

Example 73-((4-((4-(3-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2,5,8,11-tetraoxatridecan-13-yl)benzamide

Et₃N (35 μL, 0.251 mmol) was added to a stirred solution of IntermediateA1* (400 mg, 1.144 mmol) and Intermediate B3 (670 mg, 1.144 mmol) inisopropyl acetate (30 mL). The mixture was heated at 50° C. for 8 h thencooled to rt and stirred overnight during which time a precipitate wasformed. The solid was filtered off and washed with isopropyl acetate (15mL) then dried to constant weight. The product was recrystallised inMeCN (40 mL) and the resulting white solid isolated by filtrationwashing with further MeCN. The crude product was purified bychromatography on silica gel (12 g column, 1-5% MeOH in DCM) to affordthe title compound (316 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d6) δ: 9.75 (s, 1H), 9.08 (s, 1H), 8.76 (s, 1H),8.43-8.47 (m, 2H), 8.06-8.08 (m, 2H), 7.94 (d, 1H), 7.87 (s, 1H), 7.83(d, 1H), 7.63 (t, 1H), 7.57 (t, 1H), 7.37-7.48 (m, 6H), 6.56 (d, 1H),6.42 (s, 1H), 4.10 (s, 1H), 3.46-3.53 (m, 12H), 3.36-3.42 (m, 4H), 3.21(s, 3H), 2.41 (s, 3H), 1.30 (s, 9H).

LCMS m/z 841 (M+H)⁺ (ES⁺); 421 (M+2H)²⁺ (ES⁺)

Example 83-((4-((4-(3-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-methoxyethoxy)ethyl)benzamide

TEA (40.0 μl, 0.287 mmol) was added to a stirred solution ofIntermediate A1* (525 mg, 1.502 mmol) and Intermediate B4 (700 mg, 1.407mmol) in isopropyl acetate (20 mL). The mixture was heated at 50° C. for18 h. The resulting precipitate was collected by filtration and washedwith ethyl acetate (2×25 mL). The solid was recrystallised inacetonitrile to yield the title compound (571 mg) as a white solid.

¹H NMR (400 MHz, DMSO-d6) δ 9.75 (s, 1H), 9.08 (s, 1H), 8.76 (s, 1H),8.50-8.40 (m, 2H), 8.12-8.04 (m, 2H), 7.94 (d, 1H), 7.90-7.78 (m, 2H),7.67-7.60 (m, 1H), 7.60-7.54 (m, 1H), 7.52-7.32 (m, 6H), 6.55 (d, 1H),6.42 (s, 1H), 4.10 (s, 1H), 3.57-3.47 (m, 4H), 3.47-3.35 (m, 4H), 3.23(s, 3H), 2.41 (s, 3H), 1.30 (s, 9H).

LCMS m/z 753 (M+H)⁺ (ES⁺); 751 (M−H)⁻ (ES⁻)

Example 93-((4-((4-(3-(3-(tert-Butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide

Intermediate A5* (80 mg, 0.211 mmol), Intermediate B2 (114 mg, 0.211mmol) and Et₃N (10.00 μL, 0.072 mmol) were heated to 60° C. (block temp)in iPrOAc (3 mL) for 2 h. The resulting precipitate was collected byfiltration then recrystallised in acetonitrile to yield the titlecompound (63 mg) as a white solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.75 (s, 1H), 9.11 (s, 1H), 8.66 (s, 1H),8.45 (dd, 1H), 8.44 (d, 1H), 8.11-8.04 (m, 2H), 7.96 (d, 1H), 7.87 (brs, 1H), 7.82 (dd, 1H), 7.63 (ddd, 1H), 7.57 (ddd, 1H), 7.45 (dd, 1H),7.43 (d, 1H), 7.37-7.31 (m, 2H), 6.91-6.83 (m, 2H), 6.55 (d, 1H), 6.38(s, 1H), 4.10 (s, 1H), 3.56-3.46 (m, 8H), 3.42-3.35 (m, 4H), 3.21 (s,3H), 2.99 (s, 6H), 1.29 (s, 9H).

LCMS m/z 826 (M+H)⁺ (ES⁺)

Example 103-((4-((4-(3-(3-(tert-Butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide

Intermediate A5* (80 mg, 0.211 mmol), Intermediate B1 (116 mg, 0.211mmol) and Et₃N (10.00 μL, 0.072 mmol) were heated to 60° C. (block temp)in iPrOAc (3 mL) for 2 h. The mixture was concentrated under reducedpressure and the residue was purified by chromatography on the Companion(40 g column, 0-5% MeOH/DCM) to afford a gum. The gum was purified bypreparative HPLC (Gilson, Acidic (0.1% Formic acid), Acidic, WatersX-Select Prep-C18, 5 μm, 19×50 mm column, 25-75% MeCN in Water).Fractions containing product were combined, concentrated under reducedpressure then redissolved in ethyl acetate (50 mL). The organic solutionwas washed with saturated NaHCO₃ solution (50 mL), saturated brine (50mL), dried (MgSO₄) and concentrated under reduced pressure. Theresulting foam was slurried in diethyl ether (5 mL) overnight thencollected by filtration to afford the title compound (75 mg) as a whitesolid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.16 (s, 1H), 9.06 (s, 1H), 8.67 (s, 1H),8.38-8.30 (m, 1H), 8.14-8.05 (m, 2H), 8.00 (d, 1H), 7.85 (d, 1H),7.69-7.62 (ddd, 1H), 7.62-7.55 (m, 2H), 7.52-7.48 (m, 1H), 7.39-7.31 (m,3H), 6.91-6.84 (m, 3H), 6.59-6.53 (dd, 1H), 6.38 (s, 1H), 6.13 (d, 1H),3.75 (s, 3H), 3.56-3.47 (m, 8H), 3.43-3.35 (m, 4H), 3.21 (s, 3H), 2.99(s, 6H), 1.28 (s, 9H).

LCMS m/z 831 (M+H)⁺ (ES⁺); 829 (M−H)⁻ (ES⁻)

Example 113-((4-((4-(3-(3-(tert-Butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)-ethyl)benzamide

Intermediate A5* (80 mg, 0.211 mmol), Intermediate B5 (114 mg, 0.211mmol) and Et₃N (10.00 μL, 0.072 mmol) were heated to 60° C. (block temp)in iPrOAc (3 mL) for 2 h. The mixture was concentrated under reducedpressure and the residue was purified by chromatography on the Companion(40 g column, 0-5% MeOH/DCM) to afford a gum. The gum was purified bypreparative HPLC (Gilson, Acidic (0.1% Formic acid), Acidic, WatersX-Select Prep-C18, 5 μm, 19×50 mm column, 25-75% MeCN in Water).Fractions containing product were combined, concentrated under reducedpressure then redissolved in ethyl acetate (50 mL). The organic solutionwas washed with saturated NaHCO₃ carbonate solution (50 mL), saturatedbrine (50 mL), dried (MgSO₄) and concentrated under reduced pressure.The resulting foam was slurried in diethyl ether (5 mL) overnight thencollected by filtration to afford the title compound (93 mg) as a whitesolid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.21 (s, 1H), 9.17 (s, 1H), 8.67 (s, 1H),8.47 (dd, 1H), 8.14 (d, 1H), 8.13-8.07 (m, 2H), 8.00 (d, 1H), 7.93 (dd,1H), 7.85 (d, 1H), 7.66 (ddd, 1H), 7.59 (ddd, 1H), 7.42 (dd, 1H), 7.38(d, 1H), 7.36-7.31 (ddd, 2H), 6.91-6.84 (ddd, 2H), 6.61 (dd, 1H), 6.38(s, 1H), 6.12 (d, 1H), 4.19 (s, 1H), 3.58-3.46 (m, 8H), 3.43-3.35 (m,4H), 3.21 (s, 3H), 2.99 (s, 6H), 1.29 (s, 9H).

LCMS m/z 825 (M+H)⁺ (ES⁺); 823 (M−H)⁻ (ES⁻)

Example 123-((4-((4-(3-(3-(tert-Butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)-5,6,7,8-tetrahydronaphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide

A mixture of Intermediate C4(H) (250 mg, 0.471 mmol), Intermediate D2(288 mg, 0.942 mmol) and pTSA monohydrate (20 mg, 0.105 mmol) in THF (8mL) was heated at 60° C. for 6 h. The mixture was partitioned betweenEtOAc (60 mL) and aq 1M HCl (40 mL), the organic layer washed with brine(20 mL), dried (MgSO₄), filtered and evaporated under reduced pressure.The crude product was purified by chromatography on silica gel (40 gcolumn, 0-5% MeOH/DCM) to give a solid that was triturated with MeCN toafford the title compound (136 mg) as a white solid.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.79 (s, 1H), 8.68 (s, 1H), 8.47 (t, 1H),8.38 (d, 1H), 8.14 (s, 1H), 8.09 (s, 1H), 7.95 (s, 1H), 7.59 (d, 1H),7.47 (s, 1H), 7.42 (d, 2H), 7.35 (d, 2H), 6.97 (d, 1H), 6.40 (d, 1H),6.35 (s, 1H), 4.11 (s, 1H), 3.55-3.48 (m, 8H), 3.43-3.37 (m, 4H), 3.22(s, 3H), 2.39 (s, 3H), 1.75-1.60 (m, 4H), 1.28 (s, 9H). (4H under DMSO)

LCMS m/z 801 (M+H)⁺ (ES⁺)

Example 133-((4-((4-(3-(3-(tert-Butyl)-1-(2,4-dimethoxyphenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-benzamide

DPPA (160 μL, 0.742 mmol) was added to a stirred solution ofIntermediate M2 (210 mg, 0.690 mmol) and triethylamine (240 μL, 1.725mmol) in DMF (3 mL). The reaction was stirred at rt for 1 h beforeaddition of Intermediate B2 (350 mg, 0.646 mmol) and heating to 100° C.for 2 h. The reaction mixture was cooled and partitioned between EtOAc(20 mL) and 20% w/w NaCl soln. (40 mL). The organics were separated,dried (MgSO₄), filtered and evaporated to a brown gum. The crude productwas purified by chromatography on silica gel (40 g column, 2% MeOH:DCMto 8%) to afford a yellow gum which was stirred in MeCN overnight. Theresulting precipitate was filtered off and washed with MeCN (2 mL) toafford the title compound (240 mg) as a beige solid.

¹H NMR (400 MHz, DMSO-d6) δ 9.75 (s, 1H), 9.13 (s, 1H), 8.58-8.36 (m,3H), 8.10-8.01 (m, 2H), 7.98 (d, 1H), 7.90-7.85 (m, 1H), 7.83 (d, 1H),7.69-7.60 (m, 1H), 7.60-7.52 (m, 1H), 7.48-7.40 (m, 2H), 7.32 (d, 1H),6.84 (d, 1H), 6.71 (dd, 1H), 6.55 (d, 1H), 6.37 (s, 1H), 4.08 (s, 1H),3.88 (s, 3H), 3.84 (s, 3H), 3.55-3.46 (m, 8H), 3.39 (m, 4H), 3.21 (s,3H), 1.27 (s, 9H).

LCMS m/z 843 (M+H)⁺ (ES⁺)

Example 143-((4-((4-(3-(3-(tert-Butyl)-1-(3-((dimethylphosphoryl)methyl)phenyl)-1H-pyrazol-5-yl)-ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide

DPPA (80 μL, 0.371 mmol) was added to a solution of Intermediate M5 (110mg, 0.329 mmol) and triethylamine (120 μL, 0.861 mmol) in DMF (2 mL).The reaction was stirred at rt for 1 h before addition of IntermediateB2 (170 mg, 0.314 mmol) and heating at 100° C. (block temperature) for 2h. The reaction mixture was cooled and partitioned between EtOAc (20 mL)and 20% w/w NaCl soln. (40 mL). The organics were separated, dried(MgSO₄), filtered and evaporated to a brown gum. The crude product waspurified by chromatography on silica gel (40 g column, 2% MeOH:DCM to8%) to afford a beige solid which was recrystallised from MeCN (3 mL) toafford the title compound (80 mg) as a colourless solid.

¹H NMR (400 MHz, DMSO-d6) δ 9.75 (s, 1H), 9.42 (s, 1H), 8.95 (s, 1H),8.52-8.36 (m, 2H), 8.21 (d, 1H), 8.07 (s, 1H), 8.00 (d, 1H), 7.92-7.77(m, 2H), 7.70-7.51 (m, 4H), 7.52-7.38 (m, 3H), 7.32 (d, 1H), 6.56 (d,1H), 6.51 (s, 1H), 4.10 (s, 1H), 3.56-3.45 (m, 8H), 3.44-3.35 (m, 6H),3.21 (s, 3H), 1.47 (d, 6H), 1.31 (s, 9H).

LCMS m/z 873 (M+H)⁺ (ES⁺)

Example 153-((4-((4-(3-(3-(tert-Butyl)-1-(3-((dimethylphosphoryl)methyl)phenyl)-1H-pyrazol-5-yl)-ureido)naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide

DPPA (80 μL, 0.371 mmol) was added to a solution of Intermediate M5 (112mg, 0.335 mmol) and triethylamine (120 μL, 0.861 mmol) in DMF (2 mL).The reaction was stirred at rt for 1 h before addition of IntermediateB1 (170 mg, 0.311 mmol) and heating at 100° C. (block temperature) for 2h. The reaction mixture was cooled and partitioned between EtOAc (20 mL)and 20% w/w NaCl soln. (40 mL). The organics were separated, dried(MgSO₄), filtered and evaporated to a brown gum. The crude product waspurified by chromatography on silica gel (40 g column, 2% MeOH:DCM to8%) to afford a beige solid which was recrystallised from MeCN (3 ml) toafford the title compound (70 mg) as a tan solid.

¹H NMR (400 MHz, DMSO-d6) δ 9.48 (s, 1H), 9.07 (s, 1H), 8.96 (s, 1H),8.34 (t, 1H), 8.24 (d, 1H), 8.11 (d, 1H), 8.03 (d, 1H), 7.86 (d, 1H),7.71-7.63 (m, 1H), 7.63-7.44 (m, 6H), 7.36 (d, 1H), 7.35-7.29 (m, 1H),6.89 (dd, 1H), 6.57 (dd, 1H), 6.51 (s, 1H), 6.12 (d, 1H), 3.74 (s, 3H),3.57-3.46 (m, 8H), 3.43-3.35 (m, 6H), 3.21 (s, 3H), 1.47 (d, 6H), 1.31(s, 9H).

LCMS m/z 878 (M+H)⁺ (ES⁺)

Example 163-((4-((4-(3-(3-(tert-Butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide

Intermediate A5* (70 mg, 0.185 mmol), Intermediate B6 (101 mg, 0.185mmol) and Et₃N (10 μL, 0.072 mmol) were heated to 60° C. (blocktemperature) in isopropyl acetate (6 mL) and stirred for 2 h. Thecooled, gelatinous mixture was diluted with acetonitrile (6 mL) and theresulting solid was collected by filtration to yield a white solid. Thesolid was purified by preparative HPLC (Gilson, Acidic (0.1% Formicacid), Acidic, Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 5-95%MeCN in Water). Colourless needles formed in the fractions over 72 h,which were collected by filtration to afford the title compound (63 mg)as a colourless crystalline solid.

¹H NMR (400 MHz, DMSO-d6) δ: 9.61 (s, 1H), 9.15 (br s, 1H), 8.71 (br s,1H), 8.41 (d, 1H), 8.32 (dd, 1H), 8.08 (d, 1H), 7.98 (d, 1H), 7.82 (d,1H), 7.62 (ddd, 1H), 7.59-7.53 (m, 2H), 7.41 (d, 1H), 7.38-7.30 (m, 3H),6.91-6.83 (m, 3H), 6.53 (d, 1H), 6.38 (s, 1H), 3.59-3.55 (m, 2H),3.54-3.46 (m, 8H), 3.41-3.37 (m, 2H), 3.20 (s, 3H), 2.98 (s, 6H), 2.08(s, 3H), 1.28 (s, 9H).

LCMS m/z 832 (M+H)⁺ (ES⁺); 830 (M−H)⁻ (ES⁻)

Example 173-((4-((4-(3-(3-(tert-Butyl)-1-(4-methoxy-2-methylphenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide

The title compound was prepared by methods analogous to those describedabove.

¹H NMR (DMSO-d6) 400 MHz, δ: 9.74 (s, 1H), 9.09 (s, 1H), 8.58 (s, 1H),8.43-8.46 (m, 2H), 8.07 (s, 1H), 8.02 (d, 1H), 7.95 (d, 1H), 7.87 (s,1H), 7.81 (d, 1H), 7.53-7.63 (m, 2H), 7.44 (s, 1H), 7.42 (d, 1H), 7.34(d, 1H), 7.04 (d, 1H), 6.96 (dd, 1H), 6.55 (d, 1H), 6.38 (s, 1H), 4.09(s, 1H), 3.85 (s, 3H), 3.48-3.51 (m, 8H), 3.37-3.41 (m, 4H), 3.21 (s,3H), 2.02 (s, 3H), 1.29 (s, 9H).

LCMS m/z 827 (M+H)⁺ (ES⁺)

Biological Testing Experimental Methods Enzyme Binding Assays(Kinomescan)

Kinase enzyme binding activities of compounds disclosed herein may bedetermined using a proprietary assay which measures active site-directedcompetition binding to an immobilized ligand (Fabian, M. A. et al.,Nature Biotechnol., 2005, 23:329-336). These assays may be conducted byDiscoverX (formerly Ambit; San Diego, Calif.). The percentage inhibitionproduced by incubation with a test compound may be calculated relativeto the non-inhibited control.

Enzyme Inhibition Assays

The enzyme inhibitory activities of compounds disclosed herein aredetermined by FRET using synthetic peptides labelled with both donor andacceptor fluorophores (Z-LYTE, Invitrogen Ltd., Paisley, UK).

p38 MAPKα Enzyme Inhibition

The following two assay variants are used for determination of p38 MAPKαinhibition.

Method 1

The inhibitory activities of test compounds against the p38 MAPKαisoform (MAPK14: Invitrogen), are evaluated indirectly by determiningthe level of activation/phosphorylation of the down-stream molecule,MAPKAP-K2. The p38 MAPKα protein (80 ng/mL, 2.5 μL) is mixed with thetest compound (2.5 μL of either 4 μg/mL, 0.4 μg/mL, 0.04 μg/mL or 0.004μg/mL) for 2 hr at RT. The mix solution (2.5 μL) of the p38α inactivetarget MAPKAP-K2 (Invitrogen, 600 ng/mL) and FRET peptide (8 μM; aphosphorylation target for MAPKAP-K2) is then added and the kinasereaction is initiated by adding ATP (40 μM, 2.5 μL). The mixture isincubated for 1 hr at RT. Development reagent (protease, 5 μL) is addedfor 1 hr prior to detection in a fluorescence microplate reader(Varioskan® Flash, ThermoFisher Scientific).

Method 2

This method follows the same steps as Method 1 above, but utilises ahigher concentration of the p38 MAPKα protein (2.5 μL of 200 ng/mLprotein instead of 2.5 μL of 80 ng/mL protein) for mixing with the testcompound.

p38 MAPKγ Enzyme Inhibition

The inhibitory activities of compounds of the invention against p38MAPKγ(MAPK12: Invitrogen), are evaluated in a similar fashion to thatdescribed hereinabove. The enzyme (800 ng/mL, 2.5 μL) is incubated withthe test compound (2.5 μL at either 4 μg/mL, 0.4 μg/mL, 0.04 μg/mL, or0.004 μg/mL) for 2 hr at RT. The FRET peptides (8 μM, 2.5 μL), andappropriate ATP solution (2.5 μL, 400 μM) is then added to theenzymes/compound mixtures and incubated for 1 hr. Development reagent(protease, 5 μL) is added for 1 hr prior to detection in a fluorescencemicroplate reader (Varioskan® Flash, Thermo Scientific).

c-Src and Syk Enzyme Inhibition

The inhibitory activities of compounds of the invention against c-Srcand Syk enzymes (Invitrogen), are evaluated in a similar fashion to thatdescribed hereinabove. The relevant enzyme (3000 ng/mL or 2000 ng/mLrespectively, 2.5 μL) is incubated with the test compound (either 4μg/mL, 0.4 μg/mL, 0.04 μg/mL, or 0.004 μg/mL, 2.5 μL each) for 2 hr atRT. The FRET peptides (8 μM, 2.5 μL), and appropriate ATP solutions (2.5μL, 800 μM for c-Src, and 60 μM ATP for Syk) are then added to theenzymes/compound mixtures and incubated for 1 hr. Development reagent(protease, 5 μL) is added for 1 hr prior to detection in a fluorescencemicroplate reader (Varioskan® Flash, ThermoFisher Scientific).

GSK 3α Enzyme Inhibition

The following two assay variants are used for determination of GSK 3αinhibition.

Method 1

The inhibitory activities of compounds of the invention against the GSK3α enzyme isoform (Invitrogen), are evaluated by determining the levelof activation/phosphorylation of the target peptide. The GSK3-α protein(500 ng/mL, 2.5 μL) is mixed with the test compound (2.5 μL at either 4μg/mL, 0.4 μg/mL, 0.04 μg/mL, or 0.004 μg/mL) for 2 hr at RT. The FRETpeptide (8 μM, 2.5 μL), which is a phosphorylation target for GSK3α, andATP (40 μM, 2.5 μL) are then added to the enzyme/compound mixture andthe resulting mixture incubated for 1 hr. Development reagent (protease,5 μL) is added for 1 hr prior to detection in a fluorescence microplatereader (Varioskan® Flash, ThermoFisher Scientific).

In all cases, the site-specific protease cleaves non-phosphorylatedpeptide only and eliminates the FRET signal. Phosphorylation levels ofeach reaction are calculated using the ratio of coumarin emission(donor) over fluorescein emission (acceptor), for which high ratiosindicate high phosphorylation and low ratios indicate lowphosphorylation levels. The percentage inhibition of each reaction iscalculated relative to non-inhibited control and the 50% inhibitoryconcentration (IC₅₀ value) is then calculated from theconcentration-response curve.

Method 2

This method follows the same steps as Method 1 above, but utilises ashorter period of mixing of the test compound (105 minutes instead of 2hours) with the GSK3-α protein.

Cellular Assays

(a) LPS-induced TNFα/IL-8 Release in d-U937 Cells

U937 cells, a human monocytic cell line, are differentiated intomacrophage-type cells by incubation with PMA (100 ng/mL) for 48 to 72hr. Cells are pre-incubated with final concentrations of test compoundfor 2 hr and are then stimulated with LPS (0.1 μg/mL; from E. Coli:O111:B4, Sigma) for 4 hr. The supernatant is collected for determinationof TNFα and IL-8 concentrations by sandwich ELISA (Duo-set, R&Dsystems). The inhibition of TNFα production is calculated as apercentage of that achieved by 10 μg/mL of BIRB796 at each concentrationof test compound by comparison against vehicle control. The relative 50%effective concentration (REC₅₀) is determined from the resultantconcentration-response curve. The inhibition of IL-8 production iscalculated at each concentration of test compound by comparison withvehicle control. The 50% inhibitory concentration (IC₅₀) is determinedfrom the resultant concentration-response curve.

(b) LPS-Induced TNFα/IL-8 Release in PBMC Cells

Peripheral blood mononuclear cells (PBMCs) from healthy subjects areseparated from whole blood using a density gradient (Lymphoprep,Axis-Shield Healthcare). The PBMCs are seeded in 96 well plates andtreated with compounds at the desired concentration for 2 hours beforeaddition of 1 ng/ml LPS (Escherichia Coli 0111:B4 from Sigma Aldrich)for 24 hours under normal tissue culture conditions (37° C., 5% CO₂).The supernatant is harvested for determination of IL-8 and TNFαconcentrations by sandwich ELISA (Duo-set, R&D systems) and read on thefluorescence microplate reader (Varioskan® Flash, ThermoFisherScientific). The concentration at 50% inhibition (IC₅₀) of IL-8 and TNFαproduction is calculated from the dose response curve.

(c) IL-2 and IFN Gamma Release in CD3/CD28 Stimulated PBMC Cells

PBMCs from healthy subjects are separated from whole blood using adensity gradient (Lymphoprep, Axis-Shield Healthcare). Cells are addedto a 96 well plate pre-coated with a mixture of CD3/CD28 monoclonalantibodies (0.3 ug/ml eBioscience and 3 ug/ml BD Pharmingenrespectively). Compound at the desired concentration is then added tothe wells and the plate left for 3 days under normal tissue cultureconditions. Supernatants are harvested and IL-2 and IFN gamma releasedetermined by Sandwich ELISA (Duo-set, R&D System). The IC₅₀ isdetermined from the dose response curve.

(d) IL-1β-Induced IL-8 Release in HT29 Cells

HT29 cells, a human colon adenocarcinoma cell line, are plated in a 96well plate (24 hrs) and pre-treated with compounds at the desiredconcentration for 2 hours before addition of 5 ng/ml of IL-1β (Abcam)for 24 hours. Supernatants are harvested for IL-8 quantification bySandwich ELISA (Duo-set, R&D System). The IC₅₀ is determined from thedose response curve.

(e) LPS-Induced IL-8 and TNFα Release in Primary Macrophages

PBMCs from healthy subjects are separated from whole blood using adensity gradient (Lymphoprep, Axis-Shield Healthcare). Cells areincubated for 2 hrs and non-adherent cells removed by washing. Todifferentiate the cells to macrophages the cells are incubated with 5ng/ml of GM-CSF (Peprotech) for 7 days under normal tissue cultureconditions. Compounds are then added to the cells at the desiredconcentration for a 2 hour pre-treatment before stimulation with 10ng/ml LPS for 24 hours. Supernatants are harvested and IL-8 and TNFαrelease determined by Sandwich ELISA (Duo-set, R&D System). The IC₅₀ isdetermined from the dose response curve.

(f) Poly I:C-Induced ICAM-1 Expression in BEAS2B Cells

Poly I:C is used in these studies as a simple, RNA virus mimic. PolyI:C-Oligofectamine mixture (1 μg/mL Poly I:C, ±2% Oligofectamine, 25 μL;Invivogen Ltd., San Diego, Calif., and Invitrogen, Carlsbad, Calif.,respectively) is transfected into BEAS2B cells (human bronchialepithelial cells, ATCC). Cells are pre-incubated with finalconcentrations of test compounds for 2 hr and the level of ICAM1expression on the cell surface is determined by cell-based ELISA. At atime point 18 hr after poly I:C transfection, cells are fixed with 4%formaldehyde in PBS (100 μL) and then endogenous peroxidase is quenchedby the addition of washing buffer (100 μL, 0.05% Tween in PBS:PBS-Tween) containing 0.1% sodium azide and 1% hydrogen peroxide. Cellsare washed with wash-buffer (3×200 μL). and after blocking the wellswith 5% milk in PBS-Tween (100 μL) for 1 hr, the cells are incubatedwith anti-human ICAM-1 antibody (50 μL; Cell Signalling Technology,Danvers, Mass.) in 1% BSA PBS overnight at 4° C.

The cells are washed with PBS-Tween (3×200 μL) and incubated with thesecondary antibody (100 μL; HRP-conjugated anti-rabbit IgG, Dako Ltd.,Glostrup, Denmark). The cells are then incubated with of substrate (50μL) for 2-20 min, followed by the addition of stop solution (50 μL, 1NH₂SO₄). The ICAM-1 signal is detected by reading the absorbance at 450nm against a reference wavelength of 655 nm using a spectrophotometer.The cells are then washed with PBS-Tween (3×200 μL) and total cellnumbers in each well are determined by reading absorbance at 595 nmafter Crystal Violet staining (50 μL of a 2% solution in PBS) andelution by 1% SDS solution (100 μL) in distilled water. The measured OD450-655 readings are corrected for cell number by dividing with theOD595 reading in each well. The inhibition of ICAM-1 expression iscalculated at each concentration of test compound by comparison withvehicle control. The 50% inhibitory concentration (IC₅₀) is determinedfrom the resultant concentration-response curve.

(g) T Cell Proliferation

PBMCs from healthy subjects are separated from whole blood using adensity gradient (Lymphoprep, Axis-Shield Healthcare). The lymphocytefraction is first enriched for CD4+ T cells by negative magnetic cellsorting as per the manufacturer's instructions (Miltenyi Biotec130-091-155). Naïve CD4+ T cells are then separated using positivemagnetic selection of CD45RA+ cells using microbeads as per themanufacturer's instructions (130-045-901). Cells are plated at 2×10⁵cells per well in 100 μL RPMI/10% FBS on 96 well flat bottomed plate(Corning Costar). 25 μL of test compound are diluted to the appropriateconcentration (8× final conc.) in normal medium and added to duplicatewells on the plate to achieve a dose response range of 0.03 ng/mL-250ng/mL. DMSO is added as a negative control. Plates are allowed topre-incubate for 2 hours before stimulation with 1 μg/mL anti-CD3 (OKT3;eBioscience). After 72 h, the medium in each well is replaced with 150μL of fresh medium containing 10 μM BrdU (Roche). After 16 h, thesupernatant is removed, the plate is dried and the cells fixed by adding100 μL of fix/denature solution to each well for 20 min as per themanufacturer's instructions (Roche). Plates are washed once with PBSbefore addition of the anti-BrdU detection antibody and incubated for 90mins at room temperature. Plates are then washed gently 3× with the washbuffer supplied and developed by addition of 100 μL of substratesolution. The reaction is stopped by addition of 50 μL of 1 M H₂SO₄, andread for absorbance at 450 nm on a plate reader (Varioskan® Flash,ThermoFisher Scientific). The IC₅₀ is determined from the dose responsecurve.

(h) Human Biopsy Assay

Intestinal mucosa biopsies are obtained from the inflamed regions of thecolon of IBD patients. The biopsy material is cut into small pieces (2-3mm) and placed on steel grids in an organ culture chamber at 37° C. in a5% CO₂/95% C₂ atmosphere in serum-free media. DMSO control or testcompounds at the desired concentration are added to the tissue andincubated for 24 hr in the organ culture chamber. The supernatant isharvested for determination of IL-6, IL-8, IL-1β and TNFα levels by R&DELISA. Percentage inhibition of cytokine release by the test compoundsis calculated relative to the cytokine release determined for the DMSOcontrol (100%).

(i) Cell Mitosis Assay

PBMCs from healthy subjects are separated from whole blood (Quintiles,London, UK) using a density gradient (Histopaque®-1077, Sigma-Aldrich,Poole, UK). The PBMCs (3 million cells per sample) are subsequentlytreated with 2% PHA (Sigma-Aldrich, Poole, UK) for 48 hr, followed by a20 hr exposure to varying concentrations of test compounds. At 2 hrbefore collection, PBMCs are treated with demecolcine (0.1 μg/mL;Invitrogen, Paisley, UK) to arrest cells in metaphase. To observemitotic cells, PBMCs are permeabilized and fixed by adding Intraprep (50μL; Beckman Coulter, France), and stained with anti-phospho-histone 3(0.26 ng/L; #9701; Cell Signalling, Danvers, Mass.) and propidium iodide(1 mg/mL; Sigma-Aldrich, Poole, UK) as previously described (MuehlbauerP. A. and Schuler M. J., Mutation Research, 2003, 537:117-130).Fluorescence is observed using an ATTUNE flow cytometer (Invitrogen,Paisley, UK), gating for lymphocytes. The percentage inhibition ofmitosis is calculated for each treatment relative to vehicle (0.5% DMSO)treatment.

(j) Assessment of HRV16 induced CPE in MRC5 cells

MRC-5 cells are infected with HRV16 at an MOI of 1 in DMEM containing 5%FCS and 1.5 mM magnesium chloride, followed by incubation for 1 hr at33° C. to promote adsorption. The supernatants are aspirated, and thenfresh media added followed by incubation for 4 days. Where appropriate,cells are pre-incubated with compound or DMSO for 2 hr, and thecompounds and DMSO added again after washout of the virus.

Supernatants are aspirated and incubated with methylene blue solution(100 μL, 2% formaldehyde, 10% methanol and 0.175% Methylene Blue) for 2hr at RT. After washing, 1% SDS in distilled water (100 μL) is added toeach well, and the plates are shaken lightly for 1-2 hr prior to readingthe absorbance at 660 nm. The percentage inhibition for each well iscalculated. The IC₅₀ value is calculated from the concentration-responsecurve generated by the serial dilutions of the test compounds.

(k) In vitro RSV virus load in primary bronchial epithelial cells

Normal human bronchial epithelial cells (NHBEC) grown in 96 well platesare infected with RSV A2 (Strain A2, HPA, Salisbury, UK) at an MOI of0.001 in the LHC8 Media:RPMI-1640 (50:50) containing 15 mM magnesiumchloride and incubated for 1 hr at 37° C. for adsorption. The cells arethen washed with PBS (3×200 μL), fresh media (200 μL) is added andincubation continued for 4 days. Where appropriate, cells arepre-incubated with the compound or DMSO for 2 hr, and then added againafter washout of the virus.

The cells are fixed with 4% formaldehyde in PBS solution (50 μL) for 20min, washed with WB (3×200 μL), (washing buffer, PBS including 0.5% BSAand 0.05% Tween-20) and incubated with blocking solution (5% condensedmilk in PBS) for 1 hr. Cells are then washed with WB (3×200 μL) andincubated for 1 hr at RT with anti-RSV (2F7) F-fusion protein antibody(40 μL; mouse monoclonal, lot 798760, Cat. No. ab43812, Abcam) in 5% BSAin PBS-tween. After washing, cells are incubated with an HRP-conjugatedsecondary antibody solution (50 μL) in 5% BSA in PBS-Tween (lot00053170, Cat. No. P0447, Dako) and then TMB substrate added (50 μL;substrate reagent pack, lot 269472, Cat. No. DY999, R&D Systems, Inc.).This reaction is stopped by the addition of 2N H₂SO₄ (50 μL) and theresultant signal is determined colourimetrically (OD: 450 nm with areference wavelength of 655 nm) in a microplate reader (Varioskan®Flash, ThermoFisher Scientific). Cells are then washed and a 2.5%crystal violet solution (50 μL; lot 8656, Cat. No. PL7000, Pro-LabDiagnostics) is applied for 30 min. After washing with WB, 1% SDS indistilled water (100 μL) is added to each well, and plates are shakenlightly on the shaker for 1 hr prior to reading the absorbance at 595nm. The measured OD₄₅₀₋₆₅₅ readings are corrected to the cell number bydividing the OD₄₅₀₋₆₅₅ by the OD₅₀₅ readings. The percentage inhibitionfor each well is calculated and the IC₅₀ value is calculated from theconcentration-response curve generated from the serial dilutions ofcompound.

(l) The Effect of Test Compounds on Cell Viability: MTT Assay

Differentiated U937 cells are pre-incubated with each test compound(final concentration 1 μg/mL or 10 μg/mL in 200 μL media indicatedbelow) under two protocols: the first for 4 hr in 5% FCS RPMI1640 mediaand the second in 10% FCS RPMI1640 media for 24 h. The supernatant isreplaced with new media (200 μL) and MTT stock solution (10 μL, 5 mg/mL)is added to each well. After incubation for 1 hr the media are removed,DMSO (200 μL) is added to each well and the plates are shaken lightlyfor 1 hr prior to reading the absorbance at 550 nm. The percentage lossof cell viability is calculated for each well relative to vehicle (0.5%DMSO) treatment. Consequently an apparent increase in cell viability fordrug treatment relative to vehicle is tabulated as a negativepercentage.

(m) Accumulation of)β catenin in d-U937 Cells

U937 cells, a human monocytic cell line, are differentiated intomacrophage-type cells by incubation with PMA; (100 ng/mL) for between 48to 72 hr. The cells are then incubated with either final concentrationsof test compound or vehicle for 18 hr. The induction of β-catenin by thetest compounds is stopped by replacing the media with 4% formaldehydesolution. Endogenous peroxide activity is neutralised by incubating withquenching buffer (100 μL, 0.1% sodium azide, 1% H₂O₂ in PBS with 0.05%Tween-20) for 20 min. The cells are washed with washing buffer (200 μL;PBS containing 0.05% Tween-20) and incubated with blocking solution (200μL; 5% milk in PBS) for 1 hr, re-washed with washing buffer (200 μL) andthen incubated overnight with anti-β-catenin antibody solution (50 μL)in 1% BSA/PBS (BD, Oxford, UK).

After washing with washing buffer (3×200 μL; PBS containing 0.05%Tween-20), cells are incubated with an HRP-conjugated secondary antibodysolution (100 μL) in 1% BSA/PBS (Dako, Cambridge, UK) and the resultantsignal is determined colourimetrically (OD: 450 nm with a referencewavelength of 655 nm) using TMB substrate (50 μL; R&D Systems, Abingdon,UK). This reaction is stopped by addition of 1N H₂SO₄ solution (50 μL).Cells are then washed with washing buffer and 2% crystal violet solution(50 μL) is applied for 30 min. After washing with washing buffer (3×200μL), 1% SDS (100 μL) is added to each well and the plates are shakenlightly for 1 hr prior to measuring the absorbance at 595 nm (Varioskan®Flash, Thermo-Fisher Scientific).

The measured OD₄₅₀₋₆₅₅ readings are corrected for cell number bydividing the OD₄₅₀₋₆₅₅ by the OD₅₉₅ readings. The percentage inductionfor each well is calculated relative to vehicle, and the ratio ofinduction normalized in comparison with the induction produced by astandard control comprising of the Reference Compound(N-(4-(4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)pyridin-2-yl)-2-methoxyacetamide)(1 μg/m L) which is defined as 100%.

(n) IL-2 and IFNγ Release in CD3/CD28 Stimulated LPMC Cells from IBDPatients

Lamina propria mononuclear cells (LPMCs) are isolated and purified frominflamed IBD mucosa of surgical specimens or from normal mucosa ofsurgical specimens as follows: The mucosa is removed from the deeperlayers of the surgical specimens with a scalpel and cut in fragments 3-4mm size. The epithelium is removed by washing the tissue fragments threetimes with 1 mM EDTA (Sigma-Aldrich, Poole, UK) in HBSS (Sigma-Aldrich)with agitation using a magnetic stirrer, discarding the supernatantafter each wash. The sample is subsequently treated with type 1Acollagenase (1 mg/mL; Sigma-Aldrich) for 1 h with stirring at 37° C. Theresulting cell suspension is then filtered using a 100 μm cell strainer,washed twice, resuspended in RPMI-1640 medium (Sigma-Aldrich) containing10% fetal calf serum, 100 U/mL penicillin and 100 μg/mL streptomycin,and used for cell culture.

Freshly isolated LPMCs (2×10⁵ cells/well) are stimulated with 1 μg/mLα-CD3/α-CD28 for 48 h in the presence of either DMSO control orappropriate concentrations of compound. After 48 h, the supernatant isremoved and assayed for the presence of TNFα and IFNγ by R&D ELISA.Percentage inhibition of cytokine release by the test compounds iscalculated relative to the cytokine release determined for the DMSOcontrol (100%).

(o) Inhibition of Cytokine Release from Myofibroblasts Isolated from IBDPatients

Myofibroblasts from inflamed IBD mucosa are isolated as follows: Themucosa is dissected and discarded and 1 mm-sized mucosal samples arecultured at 37° C. in a humidified CO₂ incubator in Dulbecco's modifiedEagle's medium (DMEM, Sigma-Aldrich) supplemented with 20% FBS, 1%non-essential amino acids (Invitrogen, Paisley, UK), 100 U/mLpenicillin, 100 μg/mL streptomycin, 50 μg/mL gentamycin, and 1 μg/mLamphotericin (Sigma-Aldrich). Established colonies of myofibroblasts areseeded into 25-cm² culture flasks and cultured in DMEM supplemented with20% FBS and antibiotics to at least passage 4 to provide a sufficientquantity for use in stimulation experiments.

Subconfluent monolayers of myofibroblasts are then seeded in 12-wellplates at 3×10⁵ cells per well are starved in serum-free medium for 24 hat 37° C., 5% CO₂ before being cultured for 24 h in the presence ofeither DMSO control or appropriate concentrations of compound. After 24h the supernatant is removed and assayed for the presence of IL-8 andIL-6 by R&D ELISA. Percentage inhibition of cytokine release by the testcompounds is calculated relative to the cytokine release determined forthe DMSO control (100%).

(p) Human Neutrophil Degranulation

Neutrophils are isolated from human peripheral blood as follows:

Blood is collected by venepuncture and anti-coagulated by addition of1:1 EDTA: sterile phosphate buffered saline (PBS, no Ca+/Mg+). Dextran(3% w/v) is added (1 part dextran solution to 4 parts blood) and theblood allowed to stand for approximately 20 minutes at rt. Thesupernatant is carefully layered on a density gradient (Lymphoprep,Axis-Shield Healthcare) and centrifuged (15 mins, 2000 rpm, no brake).The supernatant is aspirated off and the cell pellet is re-suspended insterile saline (0.2%) for no longer than 60 seconds (to lysecontaminating red blood cells). 10 times volume of PBS is then added andthe cells centrifuged (5 mins, 1200 rpm). Cells are re-suspended inHBSS+ (Hanks buffered salt solution (without phenol red) containingcytochalasin B (5 μg/mL) and 1 mM CaCl₂) to achieve 5×10⁶ cells/mL.

5×10⁴ cells are added to each well of a V-bottom 96 well plate andincubated (30 mins, 37° C.) with the appropriate concentration of testcompound (0.3-1000 ng/mL) or vehicle (DMSO, 0.5% final conc).Degranulation is stimulated by addition of fMLP (final conc 1 μM) whichafter a further incubation (30 mins, 37° C.) the cells are removed bycentrifugation (5 mins, 1500 rpm) and the supernatants transferred to aflat bottom 96 well plate. An equal volume of tetramethylbenzidine (TMB)is added and after 10 mins the reaction terminated by addition of anequal volume of sulphuric acid (0.5 M) and absorbance read at 450 nm(background at 655 nm subtracted). The 50% inhibitory concentration(IC₅₀) is determined from the resultant concentration-response curve.

(q) Cell Cytotoxicity Assay

5×10⁴ TK6 cells (lymphoblastic T cell line) are added to the appropriatenumber of wells of a 96 well plate in 195 μL of media (RPMI supplementedwith 10% foetal bovine serum). 5 μL of DMSO control (final concentration0.5% v/v) or test compound (final concentration either 5 or 1 μg/mL) isadded to the wells and incubated at 37° C., 5% CO₂. After 24 hours, theplate is centrifuged at 1300 rpm for 3 minutes and the supernatantdiscarded. Cells are then resuspended in 7.5 μg/mL propidium iodide (PI)in PBS. After 15 minutes, cells are analysed by flow cytometry (BDaccuri). The % viability is calculated as the % of cells that are PInegative in the test wells normalized to the DMSO control.

In Vivo Screening: Pharmacodynamics and Anti-Inflammatory Activity

(A) LPS-Induced Neutrophil Accumulation in Mice

Non-fasted Balb/c mice are dosed by the intra tracheal route with eithervehicle, or the test substance at the indicated times (within the range2-8 hr) before stimulation of the inflammatory response by applicationof an LPS challenge. At T=0, mice are placed into an exposure chamberand exposed to LPS (7.0 mL, 0.5 mg/mL solution in PBS) for 30 min. Aftera further 8 hr the animals are anesthetized, their tracheas cannulatedand BALF extracted by infusing and then withdrawing from their lungs 1.0mL of PBS via the tracheal catheter. Total and differential white cellcounts in the BALF samples are measured using a Neubaur haemocytometer.Cytospin smears of the BALF samples are prepared by centrifugation at200 rpm for 5 min at RT and stained using a DiffQuik stain system (DadeBehring). Cells are counted using oil immersion microscopy. Data forneutrophil numbers in BAL are shown as mean±S.E.M. (standard error ofthe mean). The percentage inhibition of neutrophil accumulation iscalculated for each treatment relative to vehicle treatment.

(B) DSS-induced colitis in mice

Non-fasted, 10-12 week old, male BDF1 mice are dosed by oral gavagetwice daily with either vehicle, reference item (5-ASA) or test compoundone day before (Day −1) stimulation of the inflammatory response bytreatment with dextran sodium sulphate (DSS). On Day 0 of the study DSS(5% w/v) is administered in the drinking water followed by BID dosing ofthe vehicle (5 mL/kg), reference (100 mg/kg) or test compound (5 mg/kg)for 7 days. The drinking water with DSS is replenished every 3 days.During the study animals are weighed every day and stool observationsare made and recorded as a score, based on stool consistency. At thetime of sacrifice on Day +6 the large intestine is removed and thelength and weight are recorded. Sections of the colon are taken foreither MPO analysis to determine neutrophil infiltration or forhistopathology scoring to determine disease severity.

(C) TNBS-Induced Colitis in Mice

Non-fasted, 10-12 week old, male BDF1 mice are dosed by oral gavagetwice daily with either vehicle (5 mL/kg), reference item (Budesonide2.5 mg/kg) or test compound (1 or 5 mg/kg) one day before (Day −1)stimulation of the inflammatory response by treatment with2,4,6-trinitrobenzenesulphonic acid (TNBS) (15 mg/mL in 50% ethanol/50%saline). On Day 0 of the study TNBS (200 μL) is administeredintra-colonically via a plastic catheter with BID dosing of the vehicle,reference or test compound continuing for 2 or 4 days. During the studyanimals are weighed every day and stool observations are made andrecorded as a score, based on stool consistency. At the time ofsacrifice on Day 2 (or Day 4) the large intestine is removed and thelength and weight recorded. Sections of the colon are taken forhistopathology scoring to determine disease severity.

(D) Adoptive transfer in mice

On Study day 0, female Balb/C mice are terminated and spleens obtainedfor CD45RB^(high) cell isolation (Using SCID IBD cell Separationprotocol). Approximately 4×10⁵ cells/mL CD45RB^(high) cells are theninjected IP (100 μL/mouse) into female SCID animals. On study day 14,mice are weighed and randomized into treatment groups based on bodyweight. On Day 14 compounds are administered BID, via oral gavage, in apeanut oil vehicle at the dose levels outlined below and a dose volumeof 5 mL/kg. Treatment continues until study day 42, at which point theanimals are necropsied 4 hours after am administration. The colon lengthand weight is recorded and used as a secondary endpoint in the study asa measurement of colon oedema. The colon is then divided into sixcross-sections, four of which are used for histopathology scoring(primary endpoint) and two are homogenised for cytokine analysis.

Data shown is the % inhibition of the induction window between naïveanimals and vehicle animals, where higher inhibition implies closer tothe non-diseased, naïve, phenotype.

(E) Cigarette smoke model

A/J mice (males, 5 weeks old) are exposed to cigarette smoke (4%cigarette smoke, diluted with air) for 30 min/day for 11 days using aTobacco Smoke Inhalation Experiment System for small animals (ModelSIS-CS; Sibata Scientific Technology, Tokyo, Japan). Test substances areadministered intra-nasally (35 μL of solution in 50% DMSO/PBS) oncedaily for 3 days after the final cigarette smoke exposure. At 12 hrafter the last dosing, each of the animals is anesthetized, the tracheacannulated and bronchoalveolar lavage fluid (BALF) is collected. Thenumbers of alveolar macrophages and neutrophils are determined by FACSanalysis (EPICS® ALTRA II, Beckman Coulter, Inc., Fullerton, Calif.,USA) using anti-mouse MOMA2 antibody (macrophage) or anti-mouse 7/4antibody (neutrophil).

(F) Endotoxin-induced uveitis in rats

Male, Lewis rats (6-8 weeks old, Charles River UK Limited) are housed incages of 3 at 19-21° C. with a 12 h light/dark cycle (07:00/19:00) andfed a standard diet of rodent chow and water ad libitum. Non-fasted ratsare weighed, individually identified on the tail with a permanent markerand receive a single intravitreal administration into the right vitreoushumor (5 μL dose volume) of 100 ng/animal, i.v.t. of LPS (Escherichiacoli 0111:B4 prepared in PBS, Sigma Aldrich, UK) using a 32-gaugeneedle. Untreated rats are injected with PBS. Test compound,dexamethasone (Dex) or vehicle (20% hydroxypropyl-β-cyclodextrin, 0.1%HPMC, 0.01% Benzalconium chloride, 0.05% EDTA, 0.7% NaCl in deionisedwater) are administered by the topical route onto the right eye (10 μL)of animals 30 minutes prior to LPS, at the time of LPS administration,and 1, 2 and 4 hours post LPS administration. Before administration, thesolution or suspension to be administered is agitated for 5 minutes toensure a uniform suspension. 6 hours after LPS dosing, animals areeuthanized by overdose with pentobarbitone (i.v.). Following euthanasia,the right eye of each animal is enucleated and dissected into front(anterior) and back (posterior) sections around the lens. Each sectionis weighed and homogenised in 500 μL of sterile phosphate bufferedsaline followed by 20 minutes centrifugation at 12000 rpm at 4° C. Theresulting supernatant is divided into 3 aliquots and stored at −80° C.until subsequent cytokine analysis by R&D DuoSet ELISA.

Summary of In Vitro and In Vivo Screening Results

Studies conducted by LeadHunter Discover Services (DiscoveRxCorporation, Fremont, Calif.) using the KINOMEscan™ technologydetermined that compound of Example 2 did not have any effect on thebinding of the kinases B-Raf and B-Raf (V600e) to their standardligands.

TABLE 3 KinomeScan Selectivity score data for the Reference Compound andthe compound of Example 2 at 50 and 500 nM KinomeScan SelectivityScores/number of hits 50 nM 500 nM Compound S(35) S(10) S(1) S(35) S(10)S(1) Reference 0.174/67 0.083/32 0.018/7 0.370/143 0.272/105 0.117/45Compound Ex. 2 0.081/32 0.020/8  0.000/0 0.233/92  0.099/39  0.010/4 

The in vitro profile of the compound examples of the present invention,as determined using the protocols described above, are presented below(Tables 3a and 3b).

TABLE 3a The p38 MAPK (Method 2), c-Src, Syk and GSK3α (Method 2) EnzymeProfiles of Compound Examples Test Compound IC₅₀ Values for EnzymeInhibition (nM) Example No. p38 MAPKα c-Src Syk GSK3α 1 25 30 370 127732 52 11 50 3849 3 39 14 30 12159 4 50 19 46 9547 5 33 13 34 13006 6 4217 96 1695 7 88 18 237 2020 8 104 20 120 2890 9 98 15 35 12107 10 110 2548 12034 11 766 >1212 >1212 12122 12 NT NT NT 12485 13 370 21 123 212914 88 4 20 402 15 34 18 57 3196 16 NT 15 27 646 17 NT NT NT 12093

TABLE 3b Inhibition of cytokine release in stimulated cells (assays (a),(b), (c) and (d) above) IC₅₀ Values for Inhibition of Cytokine Release(nM) Test HT29 Compound dU937 cells PBMCs cells Example No. IL-8 TNFαIL-8 TNFα IL-2 IFNγ IL-8 1 NT NT 1.6 NT NT NT NT 2 0.5 0.5 2.0 0.7 44.22.5 3.3 3 NT NT 1.2 NT 7.5 2.0 NT 4 NT NT 1.4 NT 37.4 2.4 5.3 5 NT NT1.2 NT 52.1 1.7 NT 6 NT NT 2.3 NT 123.3 2.9 2.4 7 NT NT 2.4 NT 91.0 3.71.6 8 NT NT 1.6 NT 55.0 8.5 3.2 9 NT NT 2.9 NT NT 8.4 3.5 10 NT NT 3.3NT NT 2.7 4.8 11 NT NT 4.3 NT NT NT 6.2 12 NT NT 11.8 NT NT NT 10.9  13NT NT 4.6 NT NT NT 3.6 14 NT NT 1.5 NT NT NT 1.5 15 NT NT 1.2 NT NT NT2.0 16 NT NT 2.7 NT NT NT NT 17 NT NT 5.0 NT NT NT NT

In addition to the above:

-   -   when studied in assay (e) above, the compound of Example 2        exhibited IC₅₀ values of 2.1 and 2.8 nM for inhibition of        release of IL-8 and IL-6, respectively;    -   when studied in assay (g) above, the compound of Example 2        exhibited an IC₅₀ values of 2.7 nM for inhibition of T cell        proliferation; and    -   when studied in assay (p) above (neutrophil degranulation), the        compound of Example 2 exhibited an IC₅₀ of 42.7 nM (an average        of 3 experiments).

As illustrated in Table 4a, compounds of the examples of the presentinvention are markedly less active than the Reference Compound in assay(i) above, which measures impact on cell division (mitosis) in PBMCs.

TABLE 4a Effect of compounds of the examples on cell division in PBMCs(NT = not tested) Test compound % Inhibition of mitosis Reference at 5μg/mL compound 87.8^(a) 1 3.6 2 18.7 3 2.1 4 38.8 5 10.4 6 10.2 7 9.1 86.1 9 NT 10 NT 11 NT 12 NT 13 NT 14 NT 15 NT 16 NT 17 NT ^(a)See, forexample, the value reported in WO 2013/050757.

As illustrated in Table 4b, compounds of the examples of the presentinvention did not elicit any significant β-catenin induction whenstudied in assay (m) above. Thus, the potential of those compounds toincrease cellular concentrations of β-catenin was found to be negativein that their inductive effect at various test concentrations wassubstantially less than the effect produced by the Reference Compound at1 μg/mL.

TABLE 4b Effect of compounds of the examples on β-catenin induction (NT= not tested) % β-catenin induction Concentration of test compound Testcompound 1 μg/mL 5 μg/mL 10 μg/mL Reference compound 100 NT NT 1 −14 −13−17 2 0 2 0 3 1 6 6 4 5 12 NT 5 8 2 3 6 3 3 −1 7 0 −2 −5 8 6 3 2 9 1 2 410 4 2 2 11 NT NT NT 12 NT NT NT 13 −5 −2 −4 14 NT NT NT 15 4 11 27 16NT NT NT 17 NT NT NT

When studied in assay (q) above (cell cytotoxicity), percentage cellviability relative to DMSO control was measured as 101% at 5 μg/mL and103% at 1 μg/mL of the compound of Example 2. Thus, that compound doesnot exhibit any cytotoxicity in assay (q). By comparison, percentagecell viability relative to DMSO control was measured for the ReferenceCompound as 43% at 5 μg/mL and 49% at 1 μg/mL.

As illustrated in Tables 5a, 5b1 and 5b2 below, the compound of Example2 was also screened in human biopsy assay (h) and in vivo assay (C)above, as conducted over 2 days. Histopathology analysis revealed thatthe compound of Example 2 displayed significant activity in the in vivomodel of colonic inflammation. In particular, that compound, when dosedorally at 5 mg/kg, demonstrated marked improvements in ulcer grade andepithelial repair compared to the vehicle control. In addition, thecompound of Example 2 produced marked reduction in inflammatory cellinfiltrate in the reticular and lamina propria zones. The compound ofExample 2 also demonstrated marked anti-inflammatory effects in biopsiesfrom ulcerative colitis (UC) patients. In contrast to healthyvolunteers, intestinal mucosal biopsies from UC patients have been shownto spontaneously release pro-inflammatory cytokines in vitro (Onken, J.E. et al., J Clin Immunol, 2008, 126(3): 345-352). Addition of thecompound of Example 2 to biopsies in vitro markedly reduced IL-1β, IL-6and IL-8 release.

TABLE 5a Summary of results from studies on TNBS-induced colitis inmice. Experiment TNBS no. Treatment group n Ulcer grade LP inflammation1 Non-diseased 6 0.0 ± 0.0  0.2 ± 0.2  1 TNBS + Vehicle 24 4.4 ± 0.2#4.5 ± 0.2# 1 TNBS + Example 2 12 3.5 ± 0.4  2.9 ± 0.3* (1 mg/kg) 1TNBS + Example 2 12 3.0 ± 0.5* 2.2 ± 0.4* (5 mg/kg) #P > 0.001 comparedto non-diseased animals *p < 0.05 relative to vheicle control

TABLE 5b1 Summary of results from assays using intestinal mucosabiopsies from the inflamed regions of the colon of various patientssuffering from ulcerative colitis (a form of IBD). Cytokine release frombiopsies of UC patients (% release relative to DMSO control) TreatmentIL-1β IL-6 IL-8 group n release n release n release DMSO control 100%100% 100% Example 2 (1 μg/mL) 4 4 ± 10 6 29 ± 30 6 21 ± 21

TABLE 5b2 Summary of results from further assays using intestinal mucosabiopsies from the inflamed regions of the colon of various patientssuffering from ulcerative colitis (a form of IBD). Cytokine release frombiopsies of UC patients (% release relative to DMSO control) TreatmentTNFα IL-1β IL-6 IL-8 group n release n release n release n release DMSOcontrol 100% 100% 100% 100% Example 2 (1000 ng/mL) 2 10 ± 3 2 9 ± 5 4 6± 3 4  3 ± 0.1 Example 2 (100 ng/mL) 2 17 ± 1 2  2 ± 0.1 4 18 ± 4  4 19± 7  Example 2 (10 ng/mL) 2  27 ± 17 2 2 ± 2 4 40 ± 23 4 71 ± 30 Example2 (3 ng/mL) 2 48 ± 9 2 34 ± 4  4 34 ± 9  4 41 ± 14

As illustrated in Tables 5c and 5d below, the compound of Example 2 wasalso screened in cellular assays (n) and (o) above. In these assays, thecompound of Example 2 displayed significant inhibition of cytokines fromcells isolated from diseased (IBD) patients. Negative values reported inTable 5d are indicative of inhibition of the basal expression ofcytokines by the compound of Example 2.

TABLE 5c Summary of results from assays using LPMCs from IBD patients.Cytokine release from LPMCs of IBD patients (% release relative to DMSOcontrol) Treatment group n IFNγ release n TNFα release DMSO control 100100 Example 2 (1000 ng/mL) 1 2 1 0 Example 2 (100 ng/mL) 1 1 1 2 Example2 (10 ng/mL) 1 9 1 2 Example 2 (1 ng/mL) 1 8 1 12 Example 2 (0.3 ng/mL)1 20 1 35

TABLE 5D Summary of results from assays using LPMCs from IBD patients.Cytokine release from myofibroblasts of IBD patients (% release relativeto DMSO control) Treatment group n IL-8 release n IL-6 release DMSOcontrol 100 100 Example 2 (100 ng/mL) 2 −72 ± 34 2 −21 ± 8 Example 2 (10ng/mL) 2 −52 ± 22 2 −12 ± 4 Example 2 (1 ng/mL) 2    66 ± 147 2    82 ±105

As illustrated in Table 6 below, the compound of Example 2 was alsoscreened in the in vivo (adoptive transfer) assay (D) above.Histopathology analysis, as well as analysis of the relative inhibitionof cytokine release revealed that the compound of Example 2 alsodisplayed significant activity in this further in vivo model of colonicinflammation.

TABLE 5 Summary of results from adoptive transfer mouse model. Dose ofcompound of Example 2 (mg/kg) 5 1 0.2 0.04 % Inhibition of colon weight:length 61%* 43%# 23% 25%# % inhibition of IL-8 release 90%# 36%# 39%34%# % inhibition of overall histopathology score 46% 33%  5%  2% *P <0.05 ANOVA to vehicle #p < 0.05 T-test to vehicle

Summary of Additional Studies

Determination of Pharmacokinetic Parameters

(i) Studies in Mice

A study was conducted by Sai Life Sciences (Hinjewadi, Pune, India) toinvestigate the pharmacokinetics and total colon tissue distribution ofthe compound of Example 2 in male C57BL/6 mice following a single oraladministration.

A group of twenty one male mice were dosed with a suspension formulation(in peanut oil) of the compound of Example 2, at a dose of 5 mg/kg.Blood samples (approximately 60 μL) were collected from retro orbitalplexus such that the samples were obtained at 1, 2, 4, 6, 8, 12 and 24hr. The blood samples were collected from a set of three mice at eachtime point in labelled micro centrifuge tube containing K₂EDTA asanticoagulant. Plasma samples were separated by centrifugation at 4000rpm for 10 min of whole blood and stored below −70° C. untilbioanalysis. After collection of blood sample, animals were humanelyeuthanized by carbon dioxide asphyxiation to collect total colontissues. The colons were flushed with cold phosphate buffer saline (pH7.4) to remove contents. The total colon tissues were homogenized withcold phosphate buffer saline (pH 7.4) of twice the weight of colontissue and stored below −70° C. Total volume was three times the totalcolon tissue weights. All samples were processed for analysis by proteinprecipitation using acetonitrile and analyzed with developed LC-MS/MSmethod (LLOQ: 2.02 ng/mL in plasma and 1.01 ng/mL in colon tissue).Pharmacokinetic parameters were calculated using the non-compartmentalanalysis tool of Phoenix WinNonlin® software (version 6.3).

(II) Studies in Rats

A study was conducted by Sai Life Sciences (Hinjewadi, Pune, India) toinvestigate the pharmacokinetics, as well as plasma and total colontissue distribution of the compound of Example 2 in male Wistar ratsfollowing a single intravenous or oral administration.

30 male Wistar rats were divided into two groups: Group I (p.o.: 5mg/kg) and Group II (i.v.: 0.25 mg/kg). Animals in Group I wereadministered orally with an aqueous suspension formulation (having 2%HPMC and 0.5% Tween 80) of the compound of Example 2, at a dose of 5mg/kg. Animals in Group II were administered intravenously with asolution formulation (in 5% v/v DMSO, 7.5% w/v Solutol HS 15 and 87.5%saline (0.9% w/v NaCl)) of the compound of Example 2 at a dose of 0.25mg/kg. From each rat, blood samples (approximately 120 μL) werecollected from retro orbital plexus such that samples were obtained atpre-dose, 0.05, 0.13, 0.25, 0.5, 1, 2, 4, 8, and 24 hr (i.v.) andpre-dose, 0.5, 1, 2, 4, 6, 8, 12 and 24 hr (p.o.). Immediately aftercollection, plasma was harvested from blood by centrifugation and storedat −70° C. until analysis. Following collection of blood sample, theanimals (Group I) were humanely euthanized by carbon dioxideasphyxiation. The total colon was isolated, flushed with cold phosphatebuffer saline (pH 7.4) to remove contents and weighed. The total colontissues homogenized with ice-cold phosphate buffered saline, pH 7.4.Buffer volume to be used for homogenization was twice the weight oftissue. All the samples were stored below −70° C. until bioanalysis.Total colon tissue homogenate volume was three times. Plasma and totalcolon tissue samples were quantified by LC-MS/MS method (LLOQ in plasmaand total colon tissue=0.5 ng/mL).

(II) Studies in Beagle Dogs

A study was conducted by Sai Life Sciences (Hinjewadi, Pune, India) toinvestigate the plasma pharmacokinetics of the compound of Example 2 inmale beagle dogs following a single intravenous or oral administration.

A group of three male beagle dogs were administered orally with anaqueous suspension formulation (having 2% HPMC and 0.5% Tween 80) of thecompound of Example 2, at a dose of 1 mg/kg. A group of three malebeagle dogs were administered intravenously with a solution formulation(in 5% v/v DMSO, 7.5% w/v Solutol HS 15 and 87.5% saline (0.9% w/vNaCl)) of the compound of Example 2, at a dose of 0.05 mg/kg. Bloodsamples (approximately 1.5 mL) were collected from jugular vein suchthat the samples were obtained at pre-dose, 0.25, 0.5, 1, 2, 4, 6, 8,and 24 hr (p.o.) and pre-dose, 0.08, 0.25, 0.5, 1, 2, 4, 8, 12, 24 and32 hr (i.v.) post dose. The blood samples were collected from a set ofthree dogs at each time point in labelled micro centrifuge tubecontaining K₂EDTA as anticoagulant. Plasma samples were separated bycentrifugation at 2500 g for 10 min of whole blood and stored below −70°C. until bioanalysis. All samples were processed for analysis by proteinprecipitation using acetonitrile and analyzed with LC-MS/MS method(LLOQ=0.50 ng/mL). Pharmacokinetic parameters were calculated using thenon-compartmental analysis tool of Phoenix WinNonlin® (Version 6.3).

TABLE 7a Pharmacokinetic parameters determined from studies involvingoral administration of the compound of Example 2. Mouse Rat Dog Dose &route 5 mg/kg p.o. 5 mg/kg p.o. 1 mg/kg p.o. Bio matrix Plasma Totalcolon Plasma Total colon Plasma T_(max) (h) 1 8 4 8 — C_(max) (ng/mL) 1213,671 2.7 2,005 — AUC_(LAST) 49 117,473 4.8 15,528 — (h · ng/mL)AUC_(INF) NR 117,565 NC 15,761 — (h · ng/mL) F_(po) (%) — — 0.04 — 0NR—Not reported since the AUC_(INF) is 20% greater than AUC_(LAST).NC—Not calculated due to insufficient elimination phase.

TABLE 7b Pharmacokinetic parameters determined from studies involvingintravenous administration of the compound of Example 2. Rat Dog Dose0.25 mg/kg 0.05 mg/kg C₀ (ng/mL) 5,364 1,370 ± 1,175 AUC_(LAST) (h ·ng/mL) 682 218 ± 90  AUC_(INF) (h · ng/mL) 691 223 ± 92  T_(1/2) (h) 2.21.9 ± 0.4 CL (mL/min/kg) 6.3 4.1 ± 1.4 V_(d) (L/kg) 0.3 0.2 ± 0.1

TABLE 7c Concentrations of the compound of Example 2 determined atdifferent time points in the mouse pharmacokinetic study Meanconcentration of the compound of Example 2 (average of 3 experiments)Plasma Colon Time (hr) (ng/mL) (ng/g) 1 11.7     884 2 8.5     2807 45.1     4235 6 4.55 (n = 2) 1395 8 5.1     13671 12 0.0     7489 240.0     31.6

TABLE 7d Concentrations of the compound of Example 2 determined atdifferent time points in the rat pharmacokinetic study Meanconcentration of the compound of Example 2 (average of 3 experiments)Plasma Colon Time (hr) (ng/mL) (ng/g) Pre-dose 0.0    0.0 0.8    3.3 10.9       1.9 (n = 2) 2 0.6       4.0 (n = 2) 4 2.7 339 6 1.7 1862  80.6 2005  12 0.7 597 24 1.8   53.9

Determination of ADME Parameters

Assessment of certain in vitro ADME (absorption, distribution,metabolism, and excretion) parameters for the compound of Example 2 wasconducted by BioFocus (Saffron Walden, UK).

(i) Metabolic Stability

Hepatic Microsomal Stability

-   -   Microsomal stability assays were performed with incubations of        test compounds at 0.1 μM (n=2, final DMSO concentration 0.25%),        and carried out using pooled human, dog, rat and Cynomolgus        macaque hepatic microsomes from Xenotech (Lots 1210153, 0810143        and 1110042, respectively) at 0.25 mg protein/mL in the presence        of co-factor, NADPH. The incubations were performed at 37° C.        with 100 μL aliquots taken from the incubation, at 0, 2, 5, 10        and 20 minutes (and, in the case of Cynomolgus macaque hepatic        microsomes, 40 minutes) and reactions terminated by addition of        100 μL of acetonitrile containing carbamazepine as analytical        internal standard. Samples were centrifuged and the supernatant        fractions analysed by LC-MS/MS.    -   The instrument responses (peak heights) were referenced to the        zero time-point samples (as 100%) in order to determine the        percentage of compound remaining.    -   Ln plots of the % remaining, for each compound, were used to        determine the half-life for the microsomal incubations.        Half-life values were calculated from the relationship

T _(1/2)(min)=−0.693/λ

-   -   where λ was the slope of the Ln concentration vs time curve.    -   The in vitro intrinsic clearance, Cl_(int) (mL/min/kg), was        calculated and scaled to hepatic extraction ratios using the        following scaling parameters and formulae.    -   Parameters

Value Parameter Human Dog Rat Monkey Microsomal protein concentration0.25 0.25 0.25 0.25 in incubation (mg/mL) microsomes/g liver (mg) 52 7845 32 liver weight/kg body weight (g) 25 32 50 32 hepatic blood flow(mL/min/kg) 21 31 60 44

-   -   Formulae

Cl_(int)(tissue clearance)mL/min/kg=[0.693/t½(min)]×[1/microsomalprotein concentration mg/mL]×[mg microsomes/g liver]×[g liver/kg bodyweight]

Cl_(int)(hepatic clearance)mL/min/kg=hepatic bloodflow×Cl_(int)/(hepatic blood flow+Cl_(int))

Hepatic extraction ratio(Eh)=Cl_(int)(hepaticclearance)mL/min/kg/hepatic blood flow(mL/min/kg)

Cryopreserved Hepatocyte Stability

-   -   Hepatocyte stability assays were performed with incubations of        test compounds (0.1 μM initial concentration, n=2) carried out        with pooled human, dog, rat and Cynomolgus macaque cryopreserved        hepatocytes from Celsis (Lot numbers RRW, KLI and WAP,        respectively) at a cell density of 0.5 million cells/mL. The        incubations were performed at 37° C. with 100 μL samples taken        from the incubation, at 0, 10, 20, 45 and 90 minutes, and        reactions terminated by addition of 100 μL of acetonitrile        containing carbamazepine as analytical internal standard.        Samples were centrifuged and the supernatant fractions analysed        by LC-MS/MS.    -   The instrument responses (peak heights) were referenced to the        zero time-point samples (as 100%) in order to determine the        percentage of compound remaining.    -   Ln plots of the % remaining, for each compound, were used to        determine the half-life for the hepatocyte incubations.        Half-life values were calculated from the relationship

T _(1/2)(min)=−0.693/λ

-   -   where Δ was the slope of the Ln concentration vs time curve.    -   Standard compounds testosterone, midazolam and        4-methylumbelliferone are included in the assay design. These        compounds give an indication of the metabolic capacity of the        cryopreserved preparations for both Phase I and Phase II        reactions.    -   In vitro intrinsic clearance (Cl_(int)), as μL/min/million cells        was calculated by applying the following formula to the        half-life values:

Cl_(int)=0.693/T½(min)×incubation volume(μL)/million cells

-   -   The half-life values were also scaled to hepatic extraction        ratios using the scaling factors and formulae below.    -   Parameters

Value Parameter Human Dog Rat Monkey Hepatocyte concentration inincubation 0.5 0.5 0.5 0.5 (million cells/mL) Hepatocellularity (millioncells/g liver) 120 240 120 120 liver weight/kg body weight (g) 25 32 5032 hepatic blood flow (mL/min/kg) 21 31 60 44

Cl_(int)(Tissue Clearance)mL/min/kg=[0.693/t½(min)]×[1/hepatocyteconcentration (million cells/mL)]×[million cells/g liver]×[g liver/kgbody weight]

Cl_(int)(Hepatic clearance)mL/min/kg=hepatic bloodflow×Cl_(int)/(hepatic blood flow+Cl_(int))

Hepatic extraction ratio(Eh)=Cl_(int)(Hepaticclearance)mL/min/kg/hepatic blood flow(mL/min/kg)

The results catalogued in Tables 8a and 8b indicate that the compound ofExample 2 exhibits high hepatic clearance, a feature resulting in lowersystemic exposures in an in vivo setting.

TABLE 8a Summary of hepatic microsome stability tests for the compoundof Example 2 (results reported are the arithmeticmean of twoexperiments). Source of hepatic Mean intrinsic clearance Mean hepaticextraction microsomes (μL/min/mg protein) ratio (Eh) Human >554 >0.97Dog >554 >0.98 Rat 199 0.88 Cynomolgus macaque 445 0.91

TABLE 8b Summary of hepatocyte stability tests for the compound ofExample 2 (results reported are the arithmetic meanof two experiments).Mean intrinsic clearance Mean hepatic extraction Source of hepatocytes(μL/min/million cells) ratio (Eh) Human 37 0.84 Dog 46 0.92 Rat 49 0.83Cynomolgus macaque 43 0.80

(ii) Time-Dependent Inhibition of Cytochromes

CYP450 time-dependent inhibition (TDI) assays were performed with testcompound at six test concentrations, 0.062 μM to 15 μM (n=2). The testcompounds was pre-incubated for 30 minutes with pooled human hepaticmicrosomes in 0.1 M Tris buffer, pH 7.4, at 37° C. in the presence ofcofactor NADPH. A parallel series of incubations (n=2) were preparedwith no pre-incubation. Probe substrates were then added (withadditional cofactor) and further incubated for the times specified.Concentrations of probe substrates used in the incubations have beenoptimised to maintain first order reaction conditions.

Reactions were terminated with acetonitrile containing analyticalinternal standard (carbamazepine), samples then centrifuged to removemicrosomal protein and analysed using optimised LC/MS-MS conditions. TheMS data were normalized to internal standard and compared to theappropriate solvent controls to determine the amount of metaboliteformed from the probe substrate relative to the “uninhibited” controls.The results are quoted as % inhibition. These values were then plottedusing the sigmoidal dose response equation (shown below) and IC₅₀'scalculated.

Y=bottom+((top−bottom)/1+10̂((Log IC₅₀ −X)*Hill slope))

-   -   X=Log concentration    -   Y=response

IC₅₀ is quoted in μM, i.e. the point at which the inhibition is 50% ofthe control value.

Positive and negative time-dependent inhibitors were included todemonstrate the potential for specific and potent interactions under theconditions used. Variation in probe turnover between plate wells meansthat inhibition values recorded below 10-15% may not be significant.

A summary of the specific conditions are shown in the table below.

Cytochrome Microsome Probe substrate P450 conc. Conc. Incubation isoform(mg/mL) Identity (μM) Metabolite time (min) 3A4 0.25 Midazolam 7 1′-OH-15 midazolam 2C9 0.25 Diclofenac 15 1′-OH- 15 diclofenac

TABLE 9 Summary of CYP inhibition studies for the compound of Example 2(results reported are the arithmetic mean of two experiments).Cytochrome P450 0 min preincubation 30 min preincubation isoform 15 μM %Inh IC₅₀ (μM) 15 μM % Inh IC₅₀ (μM) CYP3A4 2 >15 52 7.7 CYP2C9 35 >15 37>15

Analysis of Metabolites

Studies were conducted by BioFocus (Saffron Walden, UK) to determine themetabolic fate of the compound of Example 2 following incubation withrat, dog, Cynomolgus macaque or human hepatocytes.

Separate incubations (n=3) of the compound of Example 2 (10 μM initialconcentration) or DMSO control, were performed with cryopreservedhepatocytes from each species (0.5 million cell/mL) at 37° C. for 0, 60and 90 minutes before termination of reactions and compound extractionwith acetonitrile. Sample replicates were pooled prior to analysis.

Potential metabolites were identified using time-of-flight (TOF) andtriple quadruple (TQ) mass spectrometers.

For all types of hepatocytes tested, putative metabolites of thecompound of Example 2 were observed at very low levels compared to theabundance of parent compound. The low signal intensity of somemetabolites made interpretation and structural assignment problematic.Additionally, the close relationship of the products, and theirchromatographic proximity, produced complex mass spectra from which itwas not always possible to assign fragment ions to one putativemetabolite or another. The high degree of mass resolution afforded bythe time-of-flight instrument did, however, provide confidence in theempirical formulae of the structures postulated.

A total of nine metabolites were identified in all studies. Six of thenine metabolites identified, including all of those in humanhepatocytes, had empirical formulae that were consistent with oxidativebreakdown of the polyethylene glycol (PEG) side-chain of the compound ofExample 2. The other three metabolites were observed in studies witheither dog or Cynomolgus macaque hepatocytes only.

hERG Inhibition Studies

The compound of Example 2 was tested for inhibition of the human ether ago-go (hERG) channel using IonWorks™ patch clamp electrophysiology atEssen Bioscience (Welwyn Garden City, England). Eight-pointconcentration curves were generated using serial 3-fold dilutions fromthe maximum final assay concentration (3 μM). Electrophysiologicalrecordings were made from a Chinese Hamster Lung cell line stablyexpressing the full length hERG channel. Single cell ionic currents weremeasured in the perforated patch clamp configuration (100 μg/mL)amphotericin) at room temperature (21-23° C.) using an IonWorks Quattroinstrument. The internal solution contained (mM): 140 KCl, 1 MgCl₂, 1EGTA, 20 HEPES and was buffered to pH 7.3. The external solutioncontained (mM):138 NaCl, 2.7 KCl, 0.9 CaCl₂, 0.5 MgCl₂, 8 Na₂HPO₄, 1.5KH₂PO₄, also buffered to pH 7.3. Cells were clamped at a holdingpotential of −70 mV for 30 s and then stepped to +40 mV for 1 s. Thiswas followed by a hyperpolarising step of 1 s to −30 mV to evoke thehERG tail current. This sequence was repeated 5 times at a frequency of0.25 Hz. Currents were measured from the tail step at the 5th pulse, andreferenced to the holding current. Compounds were then incubated for 6-7minutes prior to a second measurement of the hERG signal using anidentical pulse train. Eight-point concentration curves were generatedusing serial 3-fold dilutions from the maximum final assay concentration(3 μM). These studies determined that the compound of Example 2 has anIC₅₀ value for the hERG channel of greater than 3 μM (0±4% inhibition ofthe channel being observed at 3 μM concentration of the compound ofExample 2).

Diversity Profile

Studies were conducted by Cerep (Celle-Lévescault, France) toinvestigate the binding of the compound of Example 2 to a diverseselection of receptors and to investigate either the inhibition oractivation of a selection of enzymes (the “Diversity Profile” comprisinga total of 71 receptors and 26 enzymes).

When studied at a concentration of 300 nM the compound of Example 2 didnot significantly bind to any of the receptors or inhibit/activate theenzymes tested (i.e. it inhibited the control specific binding in thereceptor binding assays or enzyme assays by less than 25%, as assessedusing a suitable radioligand for each receptor or a suitable referencesubstrate for each enzyme).

Mutagenicity Assessment (Bacterial Reverse Mutation Screen)

Studies were conducted by Sequani (Ledbury, Herefordshire, UK) to assessthe compound of Example 2 in vitro for its ability to induce mutationsin two histidine dependent auxotrophic mutants of Salmonellatyphimurium, strains TA98 and TA100.

The mutation screen was conducted using the plate incorporation methodand was performed in both the presence and absence of S-9 mix (a liverpost-mitochondrial fraction derived from the livers of Aroclor 1254treated rats). The bacteria were exposed to the compound of Example 2dissolved in dimethylsulphoxide, which solvent was also used as thenegative control. The dose levels used were 0.32, 1.6, 8, 40, 200, 1000or 5000 μg/plate.

Analysable treatment levels of the compound of Example 2 were limited byinsolubility to 1000 μg/plate, as heavy precipitation observed at 5000μg/plate affected the scoring of the colonies. Precipitation was alsonoted in both strains at 1000 μg/plate in the presence and absence ofS-9 mix.

The compound of Example 2 showed no dose-related or statisticallysignificant increases in revertant colonies in either Salmonellatyphimurium strain in the presence or absence of S-9 mix. This indicatesthe absence of any mutagenic effects for the compound of Example 2 inthe Salmonella typhimurium strains studied.

Hydrolytic Stability Study

Chemical stability of the compound of Example 2 was assessed in amixture of DMSO and water (3:1) at a test compound concentration of 1mg/mL

-   -   General HPLC Procedure    -   Agilent, Waters X-Select C18, 2.5 μm, 4.6×30 mm column, 4 min        method, 5-95%    -   MeCN/water (0.1% formic acid).    -   Flow rate 2.5 ml/min.    -   Column Oven Temperature 40° C.    -   Detection 254 nm.    -   Sample Preparation    -   A 1.0 mg sample of test compound was dissolved in 750 μL of        DMSO. Water (250 μL) was added slowly, ensuring no precipitation        occurred.    -   Recording Stability    -   A 50 μL aliquot of the test solution was removed and analysed in        duplicate by 5 μL HPLC injections. The peak area for the test        compound was recorded following manual integration of the        corresponding UV trace.    -   The test solution was heated to 60° C., with stirring, and 50 μL        aliquots removed for HPLC analysis at 5 and 24 h timepoints. In        all cases, 5 μL injections were used and the samples analysed in        duplicate.    -   The peak area for the test compound was recorded at both        subsequent timepoints and the % decomposition calculated from        the % change in peak area over time.    -   Reference Compound A        (3-ethynyl-5-((4-((4-(3-(3-isopropyl-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-N-(2-morpholinoethyl)benzamide)        was included in each stability study as a control to validate        the study.

The results of the study are reported in the table below.

Test Compound Time (min) % left Reference Compound A 0 100 300 82 144036 Example 2 0 100 300 83 1440 39

Chemical stability of the compound of Example 2 (in solid form) was alsoassessed at 40° C. and 75% relative humidity. The results of the studyare reported in the table below (where chemical purity was assessedusing HPLC).

Condition 1 month, 40° C./ 3 months, 40° C./ Compound 0 months 75% RH75% RH Example 2 98.52% 96.88% 94.32%

Throughout the specification and the claims which follow, unless thecontext requires otherwise, the word ‘comprise’, and variations such as‘comprises’ and ‘comprising’, will be understood to imply the inclusionof a stated integer, step, group of integers or group of steps but notto the exclusion of any other integer, step, group of integers or groupof steps.

All patents and patent applications referred to herein are incorporatedby reference in their entirety.

The application of which this description and claims forms part may beused as a basis for priority in respect of any subsequent application.The claims of such subsequent application may be directed to any featureor combination of features described herein. They may take the form ofproduct, composition, process, or use claims and may include, by way ofexample and without limitation, the claims.

1. A compound of formula (I):

wherein: Q represents thienyl, phenyl or pyridinyl, either of which mayoptionally bear 1 to 3 substituents independently selected from,hydroxyl, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₁₋₆hydroxyalkyl, NH₂, N(H)—C₁₋₆ alkyl, N(C₁₋₆ alkyl)₂, -L-P(O)R′R″, C₁₋₆alkylene-5-10 membered heterocycle and C₀₋₃ alkylene-O—C₀₋₆alkylene-5-10 membered heterocycle; L is a direct bond or C₁₋₂ alkylene;R′ represents C₁₋₄ alkyl; R″ represents C₁₋₄ alkyl, C₃₋₆ cycloalkyl,C₁₋₄ alkoxy or hydroxy; or R′ and R″ together combine to form C₃₋₆n-alkylene, wherein one CH₂ of said n-alkylene group is optionallyreplaced by O, N(H) or N(C₁₋₄ alkyl); X represents CH or N, Y representsNR²R³; R is C₁₋₆ alkyl, C₂₋₆ alkenyl, C₁₋₆ hydroxyalkyl, C₁₋₆ haloalkyl,C₁₋₆ alkyl substituted by C₂₋₃ alkynyl, C₁₋₃ alkoxy or cyano, C₀₋₂alkylene-C₃₋₈ cycloalkyl optionally substituted with C₁₋₃ alkyl, a 4-5membered heterocycle optionally substituted with C₁₋₃ alkyl orSi(R^(1a))(R^(1b))(R^(1c)); R^(1a) and R^(1b) independently representC₁₋₄ alkyl or C₃₋₆ cycloalkyl, or R^(1a) and R^(1b) together combine toform C₂₋₆ alkylene; R^(1c) represents C₁₋₂ alkyl; R^(a) and R^(b),together with the C-atoms to which they are attached, form a fusedphenyl ring that is optionally substituted by one or more substituentsselected from C₁₋₃ alkyl, C₁₋₃ haloalkyl, cyano and halo, or one ofR^(a) and R^(b) represents H, halo, cyano, C₁₋₃ alkyl or C₁₋₃ haloalkyland the other independently represents halo, cyano, C₁₋₃ alkyl or C₁₋₃haloalkyl or R^(a) and R^(b) together represent C₃₋₅ n-alkylene, whichalkylene group is optionally substituted by one or more methylsubstituents and/or which alkylene group optionally contains one C—Cdouble bond between two C-atoms of the n-alkylene chain; R¹ is selectedfrom hydrogen, OH, halogen, CN, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₁₋₆ haloalkyl, C₀₋₃ alkylene-C₃₋₆ cycloalkyl, C₀₋₃ alkylene-O—C₁₋₃alkylene-C₃₋₆ cycloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₁₋₆hydroxyalkyl, C₀₋₃ alkylene-SO₂C₁₋₃alkyl, C₀₋₃ alkylene-SO₂NR⁴R⁵, andC₀₋₃ alkylene-NR⁶R⁷ and C₀₋₃ alkylene-NCOR⁶R⁷; one of R² and R³represents —[C₂₋₄ alkylene-O]₁₋₁₂—[C₂₋₄ alkylene]-R^(2a) and the otherof R² and R³ is selected from H, C₁₋₈ alkyl, C₀₋₆ alkylene aryl, C₀₋₆alkylene heteroaryl, —[C₂₋₄ alkylene-O]₀₋₁₂—[C₂₋₄ alkylene]-R^(2a), C₀₋₆alkylene-4-10 membered heterocycle, and C₀₋₃ alkylene-O—C₀₋₆alkylene-4-10 membered heterocycle with the proviso that when the saidheterocycle is linked through nitrogen there are at least two C-atoms inthe alkylene chain that links that nitrogen atom to the essential O atomof the substituent, wherein independently each alkyl or alkylene groupoptionally bears 1 oxo substituent, and optionally one or two carbonatoms in the alkyl or alkylene chain may each be replaced by aheteroatom selected from O, N or S(O)_(p), such that when said alkyl oralkylene comprises an amine said amino group is a tertiary amine,wherein each 4-10 membered heterocycle is optionally substituted by 1 or2 groups independently selected from halo, OH, C₁₋₆ alkyl, C₁₋₄haloalkyl, C₀₋₃ alkylene-O—C₀₋₆ alkyl, C₀₋₃ alkylene-O—C₁₋₃ haloalkyl,C₀₋₆ alkylene aryl, C₀₋₃ alkylene-O—C₀₋₃ alkylene aryl, C₀₋₆ alkyleneheteroaryl, C₀₋₃ alkylene-O—C₀₋₃ alkylene heteroaryl, C(O)C₁₋₆ alkyl,SO₂NR⁸R⁹, and C₀₋₃ alkylene-NR⁸R⁹, C₀₋₃ alkylene-NR⁸SO₂R⁹ and C₀₋₃alkylene-NR⁸C(O)R⁹; R^(2a) represents OR^(2b) or N(R^(2c)) R^(2d);R^(2b) to R^(2d) independently represent H or C₁₋₄ alkyl optionallysubstituted by one or more halo atoms, or R^(2C) and R^(2d) togetherrepresent C₃₋₆ n-alkylene, C₄₋₅ n-alkylene interrupted between C2 and C3by —O— or —N(R^(2e))— or C₆ n-alkylene interrupted between C2 and C3, orbetween C3 and C4, by —O— or —N(R^(2e))—, any of which n-alkylene groupsare optionally substituted by one or more substituents selected fromhalo, hydroxy, oxo, C₁₋₄ alkyl and C₁₋₄ alkoxy; R^(2e) represents H orC₁₋₆ alkyl optionally substituted by one or more substituents selectedfrom halo and hydroxy; R⁴ is H or C₁₋₄ alkyl; R⁵ is H or C₁₋₄ alkyl, R⁶is H or C₁₋₄ alkyl, C(O)C₁₋₃alkyl and SO₂C₁₋₃ alkyl; R⁷ is H or C₁₋₄alkyl, C(O)C₁₋₃alkyl and SO₂C₁₋₃ alkyl; R⁸ is H or C₁₋₄ alkyl, and R⁹ isH or C₁₋₄ alkyl, p is 0, 1 or 2 or a pharmaceutically acceptable saltthereof, including all stereoisomers and tautomers thereof.
 2. Acompound according to claim 1 of formula (Id1) or formula (Id2):

wherein R, R^(a), R^(b), R¹, Q and Y are as defined in claim
 1. 3. Acompound according to claim 1 of formula (If1) or formula (If2):

wherein R, R^(a), R^(b), R¹, Q and Y are as defined in claim
 1. 4. Acompound according to claim 1 of formula (Ig1) or formula (Ig2):

wherein R, R^(a), R^(b), R¹, X, Q and Y are as defined in claim
 1. 5. Acompound or salt according to claim 1, wherein R represents: C₁₋₆n-alkyl, C₃₋₆ branched alkyl, C₂₋₆ alkenyl, C₁₋₆ hydroxyalkyl, C₁₋₆haloalkyl, C₁₋₆ alkyl substituted by C₁₋₃ alkoxy or cyano, C₀₋₂alkylene-C₃₋₈ cycloalkyl optionally substituted with C₁₋₃ alkyl, or a4-5 membered heterocycle optionally substituted with C₁₋₃ alkyl.
 6. Acompound according to claim 1, wherein: R¹ represents ethynyl or OCH₃;R² represents —(CH₂CH₂O)₂₋₄CH₃; and R³ is H.
 7. A compound of formula(Ig2)

wherein: R represents isopropyl, 1-methylcyclopropyl, propen-2-yl ortert-butyl; Q represents phenyl substituted in the para position bymethyl, methoxy or dimethylamino; X represents CH or N; R¹ representsethynyl or OCH₃; Y is NR²R³; and one of R² and R³ represents—(CH₂CH₂O)₂₋₃CH₃ and the other of R² and R³ is H, or a pharmaceuticallyacceptable salt thereof, including all stereoisomers and tautomersthereof.
 8. A compound according to claim 1, wherein: Q representsphenyl substituted in the para position by methyl, methoxy ordimethylamino; and R represents isopropyl or tert-butyl.
 9. A compoundaccording to claim 1, wherein R represents tert-butyl.
 10. A compoundaccording to claim 1 selected from the group comprising or consistingof:3-ethynyl-5-((4-((4-(3-(3-isopropyl-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)-pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(4-methoxyphenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)-benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)-pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(4-methoxyphenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-methoxyethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(2,3,5,6-tetradeutero-4-(trideuteromethyl)phenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)-pyrimidin-2-yl)amino)-5-ethynyl-N-(2,5,8,11-tetraoxatridecan-13-yl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)-pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-methoxyethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)-naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)-ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)-5,6,7,8-tetrahydronaphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(2,4-dimethoxyphenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(3-((dimethylphosphoryl)methyl)phenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(3-((dimethylphosphoryl)methyl)phenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyridin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;3-((4-((4-(3-(3-(tert-butyl)-1-(4-(dimethylamino)phenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-methoxy-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide;3-((4-((4-((3-(3-(tert-butyl)-1-(4-methoxy-2-methylphenyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide,and pharmaceutically acceptable salts thereof.
 11. (canceled)
 12. Acompound according to claim 1 that is3-((4-((4-(3-(3-(tert-butyl)-1-(p-tolyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-(2-(2-methoxyethoxy)ethoxy)ethyl)benzamide,or a pharmaceutically acceptable salt thereof.
 13. A pharmaceuticalcomposition comprising a compound according to claim 1, in combinationwith one or more pharmaceutically acceptable diluents or carriers
 14. Acombination product comprising: (A) a compound according to claim 1; and(B) another therapeutic agent, wherein each of components (A) and (B) isformulated in admixture with a pharmaceutically-acceptable adjuvant,diluent or carrier.
 15. A medicament comprising a compound according toclaim
 1. 16. A method of treating a disease or condition in a subjectcomprising administering to the subject a compound according to claim 1,wherein the disease or condition is selected from the group consistingof COPD, chronic bronchitis, emphysema, asthma, pediatric asthma, cysticfibrosis, sarcoidosis, idiopathic pulmonary fibrosis, allergic rhinitis,rhinitis, sinusitis, allergic conjunctivitis, conjunctivitis,keratoconjunctivitis sicca (dry eye), glaucoma, diabetic retinopathy,macular edema, diabetic macular edema, central retinal vein occlusion(CRVO), dry and/or wet age related macular degeneration (AMD),post-operative cataract inflammation, uveitis, posterior uveitis,anterior uveitis, pan uveitis, corneal graft and limbal cell transplantrejection, gluten sensitive enteropathy (coeliac disease), eosinophilicesophagitis, intestinal graft versus host disease, Crohn's disease,ulcerative colitis, rheumatoid arthritis and osteoarthritis.
 17. Themethod of claim 16, wherein the disease or condition is selected fromthe group consisting of COPD, asthma, keratoconjunctivitis sicca (dryeye), uveitis, posterior uveitis, anterior uveitis, pan uveitis, Crohn'sdisease and ulcerative colitis.
 18. (canceled)
 19. (canceled)
 20. Aprocess for the preparation of a compound of formula (I), as defined inclaim 1, which process comprises: (a) reacting a compound of formula(II), with a compound of formula (III) wherein LG^(II) represents aleaving group and one of Z¹ and Z² is a structural fragment of formula(IV) wherein R and Q are as defined in claim 1, and the other of Z¹ andZ² is a structural fragment of formula (V) wherein R¹, R^(a), R^(b), Xand Y are as defined in claim 1; (b) reacting a compound of formula(VI), wherein R, R^(a), R^(b), Q and X are as defined in claim 1 andLG^(VI) represents a leaving group, with a compound of formula (VII),wherein R¹ and Y are as defined in claim 1; (c) reacting a compound offormula (VIII), with a compound of formula (III), wherein the compoundof formula (III) and Z¹ and Z² are as defined above; (d) reacting acompound of formula (IX), wherein Z¹ is as defined above, with anazide-forming agent, which reaction is followed, without isolation, bythermal rearrangement of the intermediate acyl azide (of formulaZ¹—C(O)—N₃) to provide, in situ, a compound of formula (VIII), whichcompound is then reacted with a compound of formula (III), as definedabove, to provide the compound of formula (I); (e) reacting a compoundof formula (X) wherein R, R¹, R^(a), R^(b), Q and X are as defined inclaim 1 and R^(X) represents H or C₁₋₄ alkyl, with a compound of formula(XI) wherein R² and R³ are as hereinbefore defined; or (f) deprotectinga protected derivative of a compound of formula (I).
 21. A compound offormula (III), as defined in claim 20, or a salt or protected derivativethereof, wherein Z² represents a structural fragment of formula (V), asdefined in claim
 20. 22. A compound of formula (VII), as defined inclaim 20, or a salt or protected derivative thereof.